University of California Berkeley
Regional Oral History Office University of California
The Bancroft Library Berkeley, California
Program in the History of the Biological Sciences and Biotechnology
Diane Pennica, Ph.D.
t-PA AND OTHER RESEARCH CONTRIBUTIONS AT GENENTECH
Interviews Conducted by
Sally Smith Hughes, Ph.D.
in 2003
Copyright 2004 by The Regents of the University of California
Since 1954 the Regional Oral History Office has been interviewing leading participants in or well-placed
witnesses to major events in the development of northern California, the West, and the nation. Oral history
is a method of collecting historical information through tape-recorded interviews between a narrator with
firsthand knowledge of historically significant events and a well-informed interviewer, with the goal of
preserving substantive additions to the historical record. The tape recording is transcribed, lightly edited
for continuity and clarity, and reviewed by the interviewee. The corrected manuscript is indexed, bound
with photographs and illustrative materials, and placed in The Bancroft Library at the University of
California, Berkeley, and in other research collections for scholarly use. Because it is primary material,
oral history is not intended to present the final, verified, or complete narrative of events. It is a spoken
account, offered by the interviewee in response to questioning, and as such it is reflective, partisan, deeply
involved, and irreplaceable.
************************************
All uses of this manuscript are covered by a legal agreement between The
Regents of the University of California and Diane Pennica dated August
6, 2003. The manuscript is thereby made available for research purposes.
All literary rights in the manuscript, including the right to publish, are
reserved to The Bancroft Library of the University of California,
Berkeley. No part of the manuscript may be quoted for publication
without the written permission of the Director of The Bancroft Library of
the University of California, Berkeley.
Requests for permission to quote for publication should be addressed to
the Regional Oral History Office, The Bancroft Library, Mail Code 6000,
University of California, Berkeley 94720-6000, and should include
identification of the specific passages to be quoted, anticipated use of the
passages, and identification of the user.
It is recommended that this oral history be cited as follows:
Diane Pennica, Ph.D., "t-PA and Other Research
Contributions at Genentech," an oral history conducted in
2003 by Sally Smith Hughes, Ph.D., Regional Oral
History Office, The Bancroft Library, University of
California, Berkeley, 2004.
Copy no.
1
Diane Pcnnica
Photo courtesy of Diane Pennica
TABLE OF CONTENTS--Diane Pennica
BIOTECHNOLOGY SERIES HISTORY by Sally Smith Hughes i
BIOTECHNOLOGY SERIES LIST jjj
INTERVIEW HISTORY v
Interview 1 : July 8, 2003 )
Tape 1 , Side A 1
Family background ]
Undergraduate work at SUNY, Fredonia 2
Graduate education at the University of Rhode Island, 1 973- 1 977 5
Postdoctoral work on vesicular stomatitus virus at Hoffmann-La Roche 6
Interviewing at Genentech 8
The academic environment at Roche 9
Tape I, Side B 9
Focus on work while at Roche 9
Early impressions of Genentech 10
First project on urokinase 1 1
Considering cloning t-PA 1 3
The importance of tenacity in science 15
Managing both t-PA and urokinase 16
RNA extraction for gamma interferon and working with Dave Goeddel 17
t-PA 18
Tape 2, Side A 18
Desire Collen 23
Being a woman in Genentech and in science 24
Out of the lab 26
The advantages of science with application 26
Interview 2: July 23, 2003 29
Tape 3, Side A 29
Tumor necrosis factor 29
Scientific publication 31
The patent process 32
Working on p53 34
Some advantages of working at Genentech 36
Tape 3, Side B 37
More on p53 37
Differences in today s scientific process 39
Staying focused on goals 42
Uromodulin 43
Tape 4, Side A 45
Cardiotropinl(CT-l) 45
Collaboration with outside researchers 50
Dealing with unsuccessful projects 52
Tape 4, Side B 52
WISP 52
Technological changes since beginning at Genentech 56
Leadership changes at Genentech 58
The ethical implications of biotechnology 59
Tape 5, Side A 59
Genentech: A pioneer 60
Defining herself as a scientist 61
Litigations 62
Most important contribution 63
APPENDICES 65
INDEX 99
BIOTECHNOLOGY SERIES HISTORY
Genesis of the Program in the History of the Biological Sciences and Biotechnology
In 1996 The Bancroft Library launched the Program in the History of the Biological Sciences and
Biotechnology. Bancroft has strong holdings in the history of the physical sciences-the papers of E.O.
Lawrence, Luis Alvarez, Edwin McMillan, and other campus figures in physics and chemistry, as well as a
number of related oral histories. Yet, although the university is located next to the greatest concentration of
biotechnology companies in the world, Bancroft had no coordinated program to document the industry or
its origins in academic biology.
When Charles Faulhaber arrived in 1995 as Bancroft s director, he agreed on the need to establish a
Bancroft program to capture and preserve the collective memory and papers of university and corporate
scientists and the pioneers who created the biotechnology industry. Documenting and preserving the
history of a science and industry which influences virtually every field of the life sciences and generates
constant public interest and controversy is vital for a proper understanding of science and business in the
late twentieth and early twenty-first centuries.
The Bancroft Library is the ideal location to carry out this historical endeavor. It offers the combination of
experienced oral history and archival personnel and technical resources to execute a coordinated oral
history and archival program. It has an established oral history series in the biological sciences, an archival
division called the History of Science and Technology Program, and the expertise to develop
comprehensive records management plans to safeguard the archives of individuals and businesses making
significant contributions to molecular biology and biotechnology. It also has longstanding cooperative
arrangements with UC San Francisco and Stanford University, the other research universities in the San
Francisco Bay Area.
In April 1996, Daniel E. Koshland, Jr. provided seed money for a center at The Bancroft Library for
historical research on the biological sciences and biotechnology. And then, in early 2001, the Program in
the History of the Biological Sciences and Biotechnology was given great impetus by Genentech s
generous pledge to support documentation of the biotechnology industry.
Thanks to these generous gifts, Bancroft has been building an integrated collection of research materials-
oral history transcripts, personal papers, and archival collections-related to the history of the biological
sciences and biotechnology in university and industry settings. A board composed of distinguished figures in
academia and industry advises on the direction of the oral history and archival components. The Program s
initial concentration is on the San Francisco Bay Area and northern California. But its ultimate aim is to
document the growth of molecular biology as an independent field of the life sciences, and the subsequent
revolution which established biotechnology as a key contribution of American science and industry.
Oral History Process
The oral history methodology used in this program is that of the Regional Oral History Office, founded in
1954 and producer of over 2,000 oral histories. The method consists of research in primary and secondary
sources; systematic recorded interviews; transcription, light editing by the interviewer, and review and
approval by the interviewee; library deposition of bound volumes of transcripts with table of contents,
introduction, interview history, and index; cataloging in UC Berkeley and national online library networks;
and publicity through ROHO news releases and announcements in scientific, medical, and historical
journals and newsletters and via the ROHO and UCSF Library Web pages.
II
Oral history as a historical technique has been faulted for its reliance on the vagaries of memory, its
distance from the events discussed, and its subjectivity. All three criticisms are valid; hence the necessity
for using oral history documents in conjunction with other sources in order to reach a reasonable historical
interpretation. Yet these acknowledged weaknesses of oral history, particularly its subjectivity, are also its
strength. Often individual perspectives provide information unobtainable through more traditional
sources. Oral history in skillful hands provides the context in which events occur the social, political,
economic, and institutional forces which shape the course of events. It also places a personal face on
history which not only enlivens past events but also helps to explain how individuals affect historical
developments.
Emerging Themes
Although the oral history program is still in its initial phase, several themes are emerging. One is
"technology transfer," the complicated process by which scientific discovery moves from the university
laboratory to industry where it contributes to the manufacture of commercial products. The oral histories
show that this trajectory is seldom a linear process, but rather is influenced by institutional and personal
relationships, financial and political climate, and so on.
Another theme is the importance of personality in the conduct of science and business. These oral histories
testify to the fact that who you are, what you have and have not achieved, whom you know, and how you
relate have repercussions for the success or failure of an enterprise, whether scientific or commercial. Oral
history is probably better than any other methodology for documenting these personal dimensions of
history. Its vivid descriptions of personalities and events not only make history vital and engaging, but also
contribute to an understanding of why circumstances occurred in the manner they did.
Molecular biology and biotechnology are fields with high scientific and commercial stakes. As one might
expect, the oral histories reveal the complex interweaving of scientific, business, social, and personal
factors shaping these fields. The expectation is that the oral histories will serve as fertile ground for
research by present and future scholars interested in any number of different aspects of this rich and
fascinating history.
Location of the Oral Histories
Copies of the oral histories are available at the Bancroft, UCSF, and UCLA libraries. They also may be
purchased at cost through the Regional Oral History Office. Some of the oral histories, with more to come,
are available on The Bancroft Library s History of the Biological Sciences and Biotechnology Website:
http://bancroft.berkeley.edu/Biotech/.
Sally Smith Hughes, Ph.D.
Historian of Science
Regional Oral History Office
The Bancroft Library
University of California, Berkeley
October 2002
1 . The three criticisms leveled at oral history also apply in many cases to other types of documentary sources.
Ill
August 2004
ORAL HISTORIES ON BIOTECHNOLOGY
Program in the History of the Biological Sciences and Biotechnology
Regional Oral History Office, The Bancroft Library
University of California, Berkeley
Paul Berg, Ph.D., A Stanford Professor s Career in Biochemistry. Science Politics, and the Biotechnology
Industry, 2000
Mary Betlach, Ph.D., Early Cloning and Recombinant DNA Technology at Herbert W. Bayer s UCSF
Laboratory, 2002
Herbert W. Boyer, Ph.D., Recombinant DNA Science at UCSF and Its Commercialization at Genentech,
2001
Roberto Crea, Ph.D., DNA Chemistry at the Dawn of Commercial Biotechnology, 2004
David V. Goeddel, Ph.D., Scientist at Genentech, CEO at Tularik, 2003
Herbert L. Heyneker, Ph.D., Molecular Geneticist at UCSF and Genentech, Entrepreneur in Biotechnology,
2004
Thomas J. Kiley, Genentech Legal Counsel and Vice President, 1976-1988, and Entrepreneur, 2002
Dennis G Kleid, Ph.D., Scientist and Patent Agent at Genentech, 2002
Arthur Komberg, M.D., Biochemistry at Stanford, Biotechnology at DNAX, 1998
Fred A. Middleton, First Chief Financial Officer at Genentech, 1978-1984, 2002
Diane Pennica, Ph.D., t-PA and Other Research Contributions at Genentech, 2004
Thomas J. Perkins, Kleiner Perkins, Venture Capital, and the Chairmanship of Genentech, 1976-1995, 2002
G Kirk Raab, CEO at Genentech. 1990-1995, 2003
George B. Rathmann, Ph.D., Chairman, CEO, and President ofAmgen, 1980-1988, 2004
Regional Characteristics of Biotechnology in the United States: Perspectives of Three Industry Insiders
(Hugh D Andrade, David Holveck, and Edward Penhoet), 2001
Niels Reimers, Stanford s Office of Technology Licensing and the Cohen/Boyer Cloning Patents, 1998
William J. Rutter, Ph.D., The Department of Biochemistry and the Molecular Approach to Biomedicine at the
University of California, San Francisco, volume I, 1998
Richard Scheller, Ph.D., Conducting Research in Academia, Directing Research at Genentech, 2002
Robert A. Swanson, Co-founder, CEO, and Chairman of Genentech, 1 976- 1 996, 2001
Daniel G Yansura. Senior Scientist at Genentech, 2002
IV
Oral histories in process:
Moshe Alafi
Brook Byers
Ronald Cape
Stanley N. Cohen
Donald Glaser
Irving Johnson
Daniel E. Koshland, Jr.
Lawrence Lasky
Arthur Levinson
Steven Rosenberg
William J. Rutter, volume II
Axel Ullrich
Mickey Urdea
Pablo Valenzuela
Keith R. Yamamoto
INTERVIEW HISTORY Diane Pennica
Diane Pennica believes that her greatest contribution to Genentech is the cloning of t-PA, an acronym for
the "clot-buster" tissue plasminogen activator, sold under the marketing name Activase. The story she tells
in this oral history is one of science, the science of isolating and cloning protein-producing genes, which
then might be made into marketable products. She has consistently in her long Genentech career she
arrived in 1980 made it her policy to stick to science because it is what she does best and because there
are others whose job it is to patent, develop, and sell. The reader will have to turn to other oral histories in
this series those with Kirk Raab and Jim Gower, for example to learn of the subsequent development
and marketing of t-PA, a product that Genentech expected to be a blockbuster.
This oral history tells of Pennica s single-minded pursuit of t-PA after a chance meeting in which she
learned of the molecule s potential for salvaging heart-attack victims through the lysing of blood clots.
But the naturally occurring substance occurred in amounts too small to be useful; the gene had to be
isolated and cloned. Genentech in 1980 was a cloning center par excellence. And Pennica was to
demonstrate that Dave Goeddel was not the only Genentech scientist with "golden hands." We learn here
of Pennica s ability and astounding tenacity fifteen hours a day, seven days a week, for two years in the
grueling and ultimately successful effort to clone the gene and isolate the complicated t-PA molecule. In
1989, she and three others received the Inventor of the Year Award from the Intellectual Property Owners
Foundation.
Although Pennica and others give pride of place to her work on t-PA, this oral history makes clear that she
has other scientific accomplishments to her credit, including work on urokinase, tumor necrosis factor,
p53, uromodulin. and cardiotropin. These are not familiar substances; Pennica introduces them in words
meaningful to the non-scientist.
She also speaks cautiously of what it was like to be one of the few women at Genentech in what, at the
start, was an aggressively male environment. She was and to some extent remains a being apart, not only
because of corporate culture but also because of her isolating focus on the doing of science and, more
recently, on the mentoring of the young people in her laboratory.
Oral History Process
Three interviews were conducted in 2003 in Pennica s office adjacent to her laboratory at Genentech.
Although she claimed nervousness, what in actuality was apparent was Pennica s intent to tell her story
chronologically and well. That she did, referring at times to her meticulously kept notebooks and
laboratory records. She reviewed the interview transcripts, editing for clarity but not changing content. By
agreement with Genentech regarding only the oral histories it supports, its legal department receives
transcripts of all interviews to review solely for current legal issues. As in all other instances to date, no
changes were requested in the Pennica transcripts.
The Regional Oral History Office was established in 1954 to augment through tape-recorded memoirs the
Library s materials on the history of California and the West. Copies of all interviews are available for
research use in The Bancroft Library and in the UCLA Department of Special Collections. The office is
under the direction of Richard Candida Smith, Director, and the administrative direction of Charles B.
VI
Faulhaber, James D. Hart Director of The Bancroft Library, University of California, Berkeley. The
catalogues of the Regional Oral History Office and many online oral histories can be accessed at http://
bancroft.berkeley.edu/ROHO/. Online information about the Program in the History of the Biological
Sciences and Biotechnology can be accessed at http://bancroft.berkeley.edu/Biotech/.
Sally Smith Hughes, Ph.D.
Historian of Science
Regional Oral History Office
The Bancroft Library
University of California, Berkeley
August 2004
Regional Oral History Office University of California
The Bancroft Library Berkeley. California 94720
BIOGRAPHICAL INFORMATION
(Please write clearly. Use black ink.)
Your full name
Date of birth 5 \ Birthplace i RgpCWtA _,
Father s full name P ft* K, J * rrV>
Occupation_ ^ftPpen Jgft. _ Birthplace Flag Po A! ft (V/t^vV V<"V3 K
Mother s full name_ MftfrVie (MAfttC. Pennica (derg<lS P
Occupation Ae^m wi s^^e A^C^f Birthplace FfrgPoJiV . K/PtJ Vofl.
Your spouse/partner_
Occupation _ 23; _ Birthplace
Your children_
Where did you grow up?
Present community_
Education TK-D Un.VeP.S . K|
Occupation(s)
Areas of expertise_ CfWce<g. Re^aRCh /
Other interests or activities
Organizations in which you are active ^A.qvi ^octft Vio,J
SIGNATURE
^<t*C- /-^iin *?-4- DATE 5"- 3/ ^ j V
INTERVIEW I: JULY 8, 2003
[Tape 1. Side A]
Hughes: Dr. Pennica, let s start way back with your grandparents on both sides. Tell me a little
bit about where they came from and what they did for a living.
Pennica: My grandparents on my mother s side were born in Sicily in a very small town called
Vallelunga, and they came over when they were both eight years old and met and got
married. My grandfather worked in the steel mill in Fredonia, New York, which is
where I grew up. My dad is a carpenter, and he built the house that I grew up in.
Hughes: Is that so?
Pennica: So we moved into that house. He built about a hundred houses in our hometown,
Fredonia, New York. My grandfather worked in the steel mill, and my grandmother had
five children. On the other side, my dad s father was a blacksmith, and my grandmother
on my dad s side had ten children eight daughters and two sons, my dad and my uncle.
It was a huge family.
Hughes: But not unusual for the time, right?
Pennica: No, not at all. My dad will be eighty in September, 2004, so, yes, they had to work to
raise money to feed the family. I grew up in an Italian household where everybody got
together on Sundays and had spaghetti and meatballs [laughs], and we had a huge
garden when I was growing up where we grew everything from corn to peas to carrots
and many other things in the garden. We always had fresh vegetables, so it s
disappointing when I go to supermarkets here.
Hughes: Yes, I can imagine.
Pennica: Because nothing tastes the same as a fresh-picked strawberry or a cherry off the tree. It
was a fun time growing up there.
Hughes: Was it an Italian community?
Pennica: Pretty much, in Fredonia. There s a lot of Italians there, and all my relatives stayed in
the same area. So we always got together with all my aunts and uncles, and we have a
huge extended family there, so it was nice.
Hughes: And are they still largely there?
Pennica: Most of them are still there, those who haven t died. They are still in the area; they
didn t travel very far. We moved into the house my dad built there when I was one year
old.
Hughes: Was carpentry considered a step up from the steel mills?
Pennica: Probably, I don t know how he and his brother decided to build homes, but they built a
lot of homes in our hometown, and my dad would show us all the homes that he d built.
His brother s wife died when she was very young of lung cancer. The two of them
would build a house, my dad s brother would move into it, they would start another one,
and he would move into that until they sold the house. If I had a second career, that s
what I would do. It s such a satisfying type of career.
Hughes: Do you go back?
Pennica: Yes, I was just there a couple weeks ago. I went there for a week. I usually go back a
couple of times a year, depending on my trips back East; so this was a nice vacation,
gorgeous weather I don t miss the hot, humid summers or the freezing cold winters,
because we get lake-effect storms, six feet of snow in the wintertime, but California
weather s perfect for me. [laughs] Hard to beat; the prices are horrible here, but it s nice
to go back and visit my hometown.
Hughes: I bet. What was the attitude about education?
Pennica: Well, my dad said, "I have four daughters, and if you want to go to college, which I
hope you all do, you have to work to pay for it." So I worked. I started working when I
was eleven years old picking berries in the fields, picking strawberries and blackberries
and currants, and I made sixty dollars for the entire summer, one summer, working from
six in the morning to six at night. I was the fastest picker and made the most money. The
entire summer, my paycheck was sixty dollars, [laughter]
I bought my first transistor radio, and boy, that was the hardest work I ve ever done in
my life. It was backbreaking, but there were people there doing that for a living, to
support their families, because they brought their kids there, and I thought, "How sad."
You know, I was doing that more for fun, and I was so thrilled that I could make that
kind of money. To me that was a huge amount of money, for a whole summer! [laughs]
Sixty dollars! So that was my first job. I then worked to support myself through college
as a bookkeeper at a Sherwin-Williams paint store, and I also worked with my mom a
bit. She worked for the Board of Cooperative Educational Services as the head
administrative assistant. It was a school for people who didn t want to go to college;
they did trade stuff, beauty culture and masonry and plumbing and things like that. I
helped her do administrative work while I was going to college. I went to college at
Fredonia State State University of New York there.
Hughes: And lived at home?
Pennica: And lived at home, right, because my parents couldn t afford to send me away to
school. I was their first daughter and I wanted to go to college, so I put myself through
school. It wasn t that expensive if you lived in the town.
Hughes: Right. Did you know what you were interested in?
Pennica: No, not at all. My dad said, "I think you should be a teacher because it s a great career
for a woman." I remember his words, he said, "You get the summers off. It s a great
thing." So I started out in college at Fredonia State as an elementary education major for
two years but took science courses on the side. Every time somebody asked me what I
was going to major in I would say, "Elementary education," and 1 would have an
uncomfortable feeling, something just wasn t right. I don t know why, and I wasn t
doing that well in the education courses you had to memorize the school system
politics in the 1800s, and 1 just didn t find that exciting. 1 think teachers are a wonderful
group of people, because without them 1 wouldn t be where I am today, but it wasn t for
me.
So I was taking science courses, and then I took a course from a man named Dr. Kevin
Fox, who taught a microbiology course at Fredonia State. He was so enthusiastic, and so
much fun. He had a spark in his eye, and he said, "Why don t you work in my lab after
class when you have breaks, and just see if you like this stuff?" So 1 was thrilled; 1 did
growth curves of bacteria, 1 did little things in the lab. Then he got me into somebody
else s lab. Dr. Irv Schmoyer, who was doing a wonderful thing. He was trying to look
for the cause of cystic fibrosis. He wanted to develop a diagnostic test for parents who
were carriers of the cystic fibrosis defect. He had a son who died when he was three
years old. 1 started working in his lab, and he had this great idea that actually worked.
He had an oyster tank in his lab, where he used the oyster gill cilia for a diagnostic
assay. The cilia are little fine hairs that move the food in for feeding, and normally these
cilia beat very synchronously like wind blowing over a wheat field. You can imagine,
they re all moving in one direction. If you take serum from a normal person, it didn t do
anything the cilia beat very regularly. However, if you took serum from either him or
his wife, because they knew they were carriers, the cilia would get all entangled, and
beat asynchronously.
Hughes: How interesting.
Pennica: So we published a paper. I was just thanked, 1 wasn t one of the authors on it, because 1
was only doing little things to help. He fractionated the serum, and he found one of the
fractions caused this asynchronous beating of the cilia in heterozygous cystic fibrosis
carriers. So that was very exciting. Between these two men, Kevin Fox and Irv
Schmoyer, they said, "Why don t you consider changing your major to science?"
Because I was always excited about going into the lab and helping out, in my junior
year, I decided to switch my major. So I crammed all the science courses in my last two
years of college I was getting A s in the science courses, and C s in the elementary
education classes because I was bored.
Hughes: [laughs] How did you happen to take the science courses on the side anyway?
Pennica: Well, I think it was probably required; I took statistics, I took economics, and one of my
teachers his name was Dr. Haitani he said, "Why don t you become an economics
major?" I was getting A s in his course, too; that was fun. But he said, and I ll never
forget his words either, he said, "You should switch your major to economics." He said,
"It s a career full of men." [laughs] 1 thought to myself, "Why should that be
important?" But he was a nice man, and he made it interesting and exciting, but I didn t
have the same sort of feeling that I did about science. I hadn t seen Kevin Fox in, oh
probably fifteen years, and just two weeks ago when I went back to Fredonia, my sister
and I went for a walk we always walk around Fredonia just to see the town we were
walking along, and I see this man in a baseball hat, and 1 say, "Good morning," just
because it s a friendly town. I got an instant recollection, and I looked back and I said,
"Kevin Fox?" And he was my old teacher that I haven t seen in probably close to
twenty years. It had been such a long time, yet he still looked the same.
Hughes: Did he remember you?
Pennica: Yes. Well, I had a baseball cap on and sunglasses, so I said, "Diane Pennica," so he gave
me a big hug, and we talked for about twenty minutes. I said, "Because of you, 1 became
a science major." I told him that he made science so much fun and so exciting. So 1
think teachers are wonderful it just wasn t a career for me.
Hughes: Were these men a little unusual? I imagine that Fredonia is rather a small town.
Pennica: Very small town.
Hughes: And the university is a small regional university?
Pennica: Yes, it s one of the SUNY schools.
Hughes: So it wasn t unusual because that research on the cilia sounds like rather a significant
discovery it wasn t unusual to have science being done of that kind of quality in a
state university?
Pennica: Well, it s hard to know because at the time, 1 didn t know what science people worked
on. To me, it was a perfect project for him because he had an emotional commitment
Hughes: Connection
Pennica: Connection to something like that, and Kevin Fox was doing some interesting research
on mice, and I learned how to handle mice from him. He was working on librium and
valium, which, he said, are supposed to calm people down, and instead he found they
were making the mice more aggressive. So 1 thought that was an interesting project all
these people were doing interesting work. I took another class from a botanist and that
was interesting to me too. So I don t know if it was cutting-edge science or not, but it
was certainly fascinating to me, so I totally switched my major thanks to these men,
who convinced me that science was exciting. If I had boring teachers, 1 probably would
have stayed in elementary education.
Hughes: Amazing thought, isn t it?
Pennica: Yeah.
Hughes: What did your parents think of your switch to science?
Pennica: Oh, they thought it was great. They could tell 1 was more excited about it. They would
ask, "What are you going to do next?" I didn t know. I didn t know what I was going to
do next.
Hughes: Had you ever thought of graduate school when you started out?
Pennica: Never, never, never. Kevin Fox and Irv Schmoyer convinced me I thought, "Oh, I ll
just get a master s degree," and they said, "Why not go for your Ph.D.?" So I
interviewed at schools, and this is again another tribute to people who can either
recognize somebody who might be excited. I got accepted at Ohio State, and
Dartmouth, and the University of Rhode Island, and everybody said, "Oh, Ohio State s
a big-time school, it s great, it s wonderful." But somebody told me that I should go
interview at these places just to see what they re like. At the time a plane ride was M>
expensive, I had no money, and I was using all my money to pay for books and college
at the time, but I thought it was a good idea. So I Hew to Ohio State, and I was shuffled
around from person to person, nobody really paid attention to me I don t even
remember what they said, but I felt disconnected. It was such a huge place, that I came
back and 1 said, "Well, it was interesting."
Then 1 went to the University of Rhode Island I decided 1 didn t want to go to
Dartmouth, 1 don t know why I flew to the University of Rhode Island, and the guy
who had an opening in his lab for graduate school, Paul Cohen, was on sabbatical in
Boston. He took time off to meet me. He was so excited about his research. He was,
again, another person with a spark, and he said, "Let me tell you about this project, and
let me tell you about this project," and I was hooked. I did not get that feeling at Ohio
State. He was just so incredible that 1 decided to get my Ph.D. from him, because he was
ready to jump out of his chair at every little experiment I did. "Oh this is so exciting!"
[laughs] He was already plotting the data in his head before 1 could even get back to my
desk. As data was coming off the scintillation counters, he was so excited.
Hughes: What was he? Was this molecular biology?
Pennica: It was microbiology.
Hughes: Microbiology
Pennica: Yeah, and we were studying viruses at the time, and actually bacteriophage, viruses that
infect bacteria. We were looking at you wanted to know what I was studying?
Hughes: Yes, yes, 1 do.
Pennica: Oh, it s so long ago, but I do remember. We were studying messenger RNA stability and
decay in T-4-infected E.coli. So when the virus infects the bacteria, it takes over the
machinery of the cell, and it makes its own RNA. We were looking at how that RNA
behaved, how long it lasted in the cell, and how quickly it got degraded. And that was
fun for me. He was just the most enthusiastic teacher. He s still doing great work now
but he was terrific, and he convinced me, "Oh, just don t stop at getting your master s,
just go on and get your Ph.D." So I did, because of this teacher encouraging me. I didn t
know what I wanted to do, but everything just fell into place. I always loved working in
the lab. I felt I was good at it, and it was really fun.
Hughes: Was there a science community at the university, or was Cohen kind of a fish out of
water?
Pennica: Well, there was a little community there that was in the biology department. I took
courses from the biophysics guy. There was another woman there who was just thinking
about recombinant DNA, because plasmids had just been discovered. One question that
she asked during my thesis oral exam that I ll never forget and that I didn t know the
answer to was, "What do you think plasmids could be useful for?" Of course, that s
[laughs] all biotechnology; I said, "I don t know, but
Hughes: What year was that?
Pennica: 1 got my Ph.D. in four years, so
Hughes: Nineteen seventy-seven.
Pennica: Seventy-seven, right; so that s before [Herbert W.] Boyer and [Stanley N.] Cohen, and
[pause] Swanson
Hughes: No, it s not before Genentech is formed in April of 1976.
Pennica: Seventy-six, right.
Hughes: So Genentech is barely off the ground. Cohen and Boyer s first paper was 1973
Pennica: Oh, okay.
Hughes: But this shows you, I mean it gives you an idea of how this, what we now recognize as
revolutionary, science takes a while to diffuse.
Pennica: Oh, exactly.
Hughes: You re not the first person to say that. I mean, it s not as though all of a sudden 1973 or
even 1974 everybody switches to recombinant DNA.
Pennica: Exactly. So we were studying RNA stability. 1 had never heard of a plasmid at this
point. I didn t know about restriction enzymes, we weren t using them. We were very
focused on what we were doing, so I didn t know about what was going on with
recombinant DNA. But this woman said, "Plasmids, could they be useful for anything?"
She must have read all of this stuff, and I had no clue about them because I just read the
papers that were focused on what I was doing, not
Hughes: Was the work that you were doing with bacteriophage and the messenger RNA, was it a
microbial/biochemical approach, would you say?
Pennica: Yes, right. And because of that, it helped me get a job here, because that s what they
were interested in. They wanted somebody who knew how to work with RNA because
that was critical. That was one of the things that helped get me hired, I believe.
Hughes: Yes. But there s a step in between
Pennica: Oh, did I miss something?
Hughes: Well, you were at [Hoffmann-La] Roche.
Pennica: Oh, right, yes, yes.
Hughes: Tell me how that happened.
Pennica: Yeah, the story of how I got to here is also an amazing story.
Hughes: Well, we ll tell that next.
Pennica: But Roche. Paul Cohen had a friend at the Roche Institute of Molecular Biology, Herb
I inns, and he was also doing similar work, but with animal viruses studying stability
and messenger RNA decay in vesicular, stomatitis virus. That s a virus that s similar to
hoof and mouth disease, [pause] So we were doing similar things, so it was a natural
progression for me to go there, leam about a slightly different system, but do similar
experiments. That was a good fit, so I went to the Roche Institute and did my postdoc
work there for two years, and that was a lot of fun. Herb Ennis was a very meek and
mild man, and we had a lot of fun together. There was a meeting which tells you how
I got here there was a meeting in London on rhabdoviruses. This family includes
rabies virus and vesicular stomatitis virus, which I was working on. A friend of mine
upstairs at the institute said he was going to go to this meeting, and he said, "Why don t
you try to go?" 1 asked my boss, and he said, "1 don t have the money to send you." I
had seen an advertisement in one of the science magazines, where they said, "If you
have some exciting research, we have some fellowships available for $500." And 1 said,
"Herb, look at this, maybe I can get some money to go to this meeting." He said, "Oh,
people never get those, don t waste your time." I didn t listen to him!
So I wrote a little paragraph on what 1 was doing. I was working on VSV stability and
decay, and we had found something very interesting. This virus, VSV, has only five
messenger RNAs, that it uses to take over the cell, and nobody up to that time had ever
separated two of them. They were of similar size and charge and molecular weight that
when you ran them out on a gel, they co-migrated together. So we devised a urea
agarose RNA gel system that 1 would run in the cold room for about eight hours. We
would pour these huge long gels, and finally, we were able to see a very, very thin
separation between these two RNAs on the gel. 1 was able to cut them out of the gel, put
them in a test tube, and figure out which RNA made which protein. This was so
exciting, because nobody had ever done this. So that was one of the first papers I
published at Roche, saying, identification of this band makes this protein in this virus.
Nobody had ever done that before. I wrote this in an abstract, because I was very
excited, and they wrote back and said, "We ll give you the money for the conference."
Herb was floored! He said, "I can t believe it." So I said, "Well, if you don t try, you
don t get!" He was a shy, meek person, but I said, "I want to try. Why not?" So, my
friend Doug Testa, who was also working on some other virus, and I went to London
together.
I didn t apply to give a talk at the meeting, but they just wanted to know what you were
working on, and why you should be given the money, and why this conference was
relevant to you. Doug said, "Diane, when you go to a meeting" this was my very first
meeting "always bring a few slides." 1 said, "Why? I m not going to talk." I was shy,
very, very shy still am. [laughs] But he said, "Bring a few slides." He said, "Well, if
you re talking to somebody and they ask about your research, you can show them your
data. I mean, you ve done this incredible thing; you ve separated these RNAs. Nobody
knows about this stuff yet" I don t think our paper had come out yet "so this is really
exciting. I said, "Okay, fine, good idea." So we went to the meeting, and he said, "I m
going to ask" I was telling him about this stuff on the plane, and he said, "This is
incredibly exciting, so I m going to ask one of the organizers if you can speak." I said,
"No, no, no, please; I haven t prepared a talk besides, I m a nervous wreck, I didn t
want to do it." He said, "Just let me ask." He went to the back, and he said, "You ve got
to let Diane speak. She s got something really relevant and pertinent to this meeting."
The guy said, "I ll give her five minutes, five slides." Now this is very unheard of, to
put a totally unknown speaker, somebody they don t even know on the program. You
normally submit abstracts, and tell them what you re going to say they didn t know
what I was going to say up there. But Doug Testa convinced me again, somebody was
pushing me to do this. And I said, "Oh God!" So 1 went back, he said, "They re
putting you at the end of the session and letting you speak for five minutes." And 1 go,
"God!" I went and put seven slides back in the projector, although he told me only five!
And 1 probably gave the best talk that I ve ever given in my life because 1 was so
nervous, so hyped-up. And in five minutes 1 was able to tell the story here s the virus,
there s five RNAs, I was able to separate them, this is what they look like separated, I
put them in a test tube, I translated them, this makes this, this makes that, boom. And
everybody was floored, and they clapped, and I got so many people coming up saying,
"That was an incredible talk," because most people talked for half an hour, and you
often get bored.
Hughes: Yes, yes.
Pennica: Five minutes is probably a good thing; you just tell the essence, and not a lot of
background. My five-minute talk was the key to my getting a job at Genentech, because
in the audience there was a guy from the Centers for Disease Control in Atlanta, Jack
Obijeski, whom you may have heard of.
Hughes: I ve heard his name, yes.
Pennica: Jack came up to me after my talk and said, "So, you think you can separate RNAs? It
looks like you re good at working with RNA." I said, "Well, I did it once, sure I can do
it." And he said, "We re working with a rabies virus can you do the same thing with
that?" And another virus, I can t remember the name of it, it just slipped my mind.
Related, again, related. But they couldn t separate the RNAs. And I said, "Sure." He
was at the CDC in Atlanta, and he sent me the RNAs, and I did the same thing,
separated them, translated them, and determined which RNA made which protein. We
published a paper. Then he said, "There s a new company that s just started out that has
been contacting me, called Genentech, and they are interested in hiring people, can I
give them your name? Because they want people who know how to work with viruses.
They re actually trying to make a vaccine for rabies, and you ve been working on it, and
you know how to work with RNA, that s what they re interested in." And I said, "Sure,
give them my name."
Hughes: Never, of course, having heard of Genentech
Pennica: Never having heard of it. I had no clue what they were even about. And they gave me a
call. So because I persisted, wrote this abstract, gave that five-minute talk, Jack and I
collaborated on the rabies project, which he told Genentech about, and they gave me a
call. Mike Ross called me and said, "We want somebody who knows how to work with
RNA, who s worked with viruses." They had me out there for an interview, and I got
hired. So, just a series of little things, and people pushing or helping along the way got
me here. I gave a seminar in Building One, this tiny little room, and people were lying
on the floor because there weren t enough seats. I told the story of separating the
viruses, and all the RNA work that I had done. I also interviewed at Chiron, and they
did not hire me.
Hughes: Isn t that interesting?
Pennica: So, 1 don t think they were doing RNA work, or that wasn t something that they were
interested in, 1 don t know why. But, Genentech offered me a job.
Hughes: Interesting. Well, before we leave Roche, say something about the Roche Institute:
What was the atmosphere? Was it closer to what you had experienced in academia, or
were you beginning to move more into a corporate sort of culture?
Pennica: No, it was academia, totally.
Hughes: It was, totally.
Pennica: Yes, because the institute was completely separate from the company. The company
seemed different; we were a little building on the hill that was very isolated, and I felt it
was a little research center, that we were doing what we wanted.
Hughes: Oh, is that so?
Pennica: 1 think so they just had a research institute that I thought was great because the people
were doing lots of exciting research. But, I don t know if any of it translated down to
what was happening at the company itself.
Hughes: Well, the fact that you weren t aware says something right there.
Pennica: Exactly. It seemed very separate. And we didn t mingle with them; we had our own
little tower. It was separate. It seemed very academic to me. And again, I just focused in
the lab, and that s all 1 did.
Hughes: Did you have any thoughts, when you got the job offer from Genentech, about, oh,
wow, this means I m leaving academia, and I m moving into the corporate world?
Pennica: No, I was so naive. Again, that s probably a good thing because people are struggling
now, oh, shall I stay in academics, should I stay in industry? I never even had that
thought for a second because 1 didn t know what Genentech was. I didn t know, [pause]
Again, I think it was probably better for me not to know what it was all about. To go all
the way across the country, and [pause] You know, I was interviewed by Herb
Heyneker, Peter Seeburg, I believe, 1 don t know if Dave Goeddel, my eventual boss, if
he interviewed me. At the time I think he was in the middle of an experiment
[Tape l,SideB]
Hughes: What about the new genetic technologies at Roche? I mean, were people using
recombinant DNA, or
Pennica: I think they were starting to but, again, I had blinders on 1 was so focused on what I
was doing, that I wasn t the type to go down the hall and say, "Hey, what are you
doing?" I probably should have because I would have been more exposed to that. I think
people were using restriction enzymes there. And eventually somebody who went to
Chiron, Mike Innis, I think he was starting to do that sort of work. But again I always
worked so intensely that I didn t have time to run around and ask people what they were
doing. I would hear their seminars. We had a Nobel Prize winner at the institute, Severe
Ochoa
Hughes: Oh, yes, I know.
Pennica: He was two doors down from my lab, and I would go in and talk to him occasionally.
Hughes: He was approachable, then?
Pennica: Oh yeah, he was a really nice man; he seemed very old [laughs] at the time. He moved
very slowly, but he was great. Everybody was nice there, and 1 had a really good time. It
was almost too short most people do postdocs for four years; I was there about a year
and a half. And that was it, but 1 did publish papers. I don t remember how many it s
on my CV. But it was a fun place to work, and Herb Ennis was a great guy to work for,
too.
Hughes: It sounds as though, if you were not even going down the hall very much, that you were
working very intensely.
Pennica: Yes, yes.
Hughes: Is that your style?
Pennica: Yes, and my boss said, "You have no clue what s going on in the rest of the company,"
because I usually don t! [laughs] So, that s a good thing and a bad thing. But I tend to
focus and not get involved in politics, and I think it s good for me.
Hughes: Can you remember your first impressions of Genentech?
Pennica: Yes. [pause] My first impressions when I was hired? Or before, as I was being
interviewed?
Hughes: Well, maybe when you were being interviewed, how did it compare with the other
institutions that you d been in? Did this group seem different in any way?
Pennica: Yes, more laid back. Chiron was very formal. There was a beautiful conference room,
lots of people in it, very formal thing. And the room I gave my seminar in at Genentech,
people were lying on the floor because it was such a tiny room. It seemed very informal,
and I became less nervous as I kept talking because they seemed so laid back. But they
asked a lot of questions, and they seemed interested. It just seemed like they were a
bunch of young people, but I didn t know what the company was about, until I talked to
them later, and they told me, "Here s what were trying to do."
Hughes: Was [Robert A.] Swanson at that first meeting?
Pennica: I don t think he was at my seminar. I don t even remember Mike Ross was there, Herb
Heyneker, Peter Seeburg was laying on the floor, I think Axel Ullrich was there, Hugh
Nile. I think Hugh Nile was interviewing at the same time, on the same day he gave a
seminar as well; I think he heard my seminar. There was a handful of people there, so
II
just the people who were involved in hiring me like Herb Heyncker, who was my first
boss here were in that little room.
\ highes: Does that mean that, at least at that stage, that the scientists pretty much hired the
scientists?
Pennica: I think so. Yes.
Hughes: And they could do that independently of Swanson?
Pennica: Well, I don t know if that was the call the letter that I got saying, "We d like to hire
you," was from Bob Swanson. I probably still have that letter. I m sure 1 do; I ve never
thrown it away. Herb Heyneker was supposed to be my boss, but I think it was up to the
scientists to decide whether the caliber of the science of the people who they were
trying to hire was good enough.
Hughes: Yes, so probably somebody or somebodies then went to Swanson and said, "This
woman is good. She fits right in, and we should hire her."
Pennica: Probably, yeah. I would hope that s what they said!
Hughes: Why Heyneker?
Pennica: Who knows what the formality was at that point. He either had a position in his lab, or
the project he was working on needed someone who knew how to work with RNA. 1
don t know why it wasn t Dave Goeddel I eventually did work for Dave but I don t
know how it was decided. Because there weren t that many people there at the time. I
could have worked for Peter Seeburg or Axel Ullrich or Dennis Kleid. Dennis may have
been at my talk, I don t know; he would probably remember if he heard my first
seminar. I don t remember if he was in the room because I didn t know these names and
faces, but 1 bet Dennis would remember. You might have met him, Dennis Kleid
you ve interviewed him, too.
Hughes: I ve interviewed him, yes, yes.
Pennica: So, he would remember that.
Hughes: And then, what were you assigned to do in your windowless lab? [laughs]
Pennica: Interesting. I was assigned to work on the very first contract project that Genentech had
with a German company called Griinenthal, and it was to clone urokinase 1 didn t
know what that was at the time a serine protease that is made in the urine that
dissolves fibrin. They were thinking of using that for potential treatment of blood clots.
On that project, we were collaborating with this company and getting protein sequence,
and we were using cells I can t remember the name of the cells human embryonic
kidney cells, I believe, to extract the RNA to see if we could clone urokinase. Bob
Swanson, because it was our first project, was very adamant about doing this well, you
know, doing a good job, because he wanted to make a good impression since they were
giving us money.
12
Hughes: Why do you call it the first project?
Pennica: It probably wasn t the first it was my first project.
Hughes: Oh, your first project, I see.
Pennica: My first project. It might have been the first contract, I don t know. You probably know
better than I do whether we had contracts with growth hormone, or somatostatin, or
insulin I don t know if we had contracts or we were just doing them.
Hughes: Certainly the insulin contract came after the cloning.
Pennica: Right.
Hughes: Genentech had to prove its stuff, that it indeed could do that.
Pennica: Griinenthal was just the opposite, I think. They wanted to clone this, and they were
giving us money before we cloned it.
Hughes: Well, and it probably does relate to that past history, 1980. See, growth hormone had
been cloned too, so probably Griinenthal knew about this small company that seemed to
be the place to go for cloning.
Pennica: Yes, yes. I m sure that that s probably true, but I thought Bob had said this was our first
contract.
Hughes: That could be, that could be.
Pennica: His office was as close as the one here to my lab, so he would be in my lab all the time
asking, "How are you doing?" and "What s going on?" and "What s new?"
Hughes: How much freedom were you given? Was this really your other than Swanson looking
over your shoulder, I mean.
Pennica: They left the experiments up to me, for sure, and I was working with another research
assistant there, Bill Holmes, who eventually got his Ph.D. not at Genentech, he got it
in Belgium and that s a different story but the two of us were working on urokinase
together. He was working on one aspect of the cloning. I was doing the RNA work and
doing similar things to what I was doing at Roche, since I had that expertise. I would
extract RNA, see if it would make in a test tube urokinase that we could detect with an
antibody. So they didn t tell you what experiments to do, I was just doing them.
Hughes: And was that all right with you? Or was that a little intimidating?
Pennica: Well, I wasn t sure where to start at this point, but I knew the steps I had to do, at least at
that point. Then, I d been at Genentech one month, and they said that they needed
somebody to go to this fibrinolysis meeting in Sweden because they had heard rumors
about somebody working on a blood-clot dissolving substance. I don t know if they
knew the name of it; at that point they might have. But Gordon Vehar said, "We need
somebody to go." Dave couldn t go, he was too busy, Herb had a family, everybody else
13
was too busy, and 1 said. "Sure, I ll go to this meeting." They said, "l ; ind out what is
known, what s going on there might be something that s better than urokinase, or as
good as, that we should know about it.". And that s where the story picks up that you had
heard, that they sent me to this meeting, and by accident I got into this private meeting
by mistake. 1 don t know if you wanted me to
Hughes: Tell the story because others don t know it.
Pennica: Right. So it was my first, no actually second now, because I d been to London already,
that was the second the first meeting was how I got to Genentech, by giving that little
five-minute talk. So this was my second international trip, and I was very nervous. 1 am
so particular about time. I worry about being late, it was instilled in me when I was
growing up, be on time, be on time, be on time. 1 always keep that in the back of my
mind. I m always early for things, and I think that s better; to this day 1 try to be early
for things because I tend to meet people who 1 never would meet otherwise. And so, 1
get off the plane and, since the meeting started the next day, I wanted to map it out; I
wanted to walk to the meeting ahead of time, just so I wasn t nervous, and try to figure
out where it was because I knew it was a little out of the way. 1 thought, if I did this I ll
be confident in the morning, because I ll know exactly how long it takes to get there,
since I mapped it out ahead of time. I start checking into the hotel, and I asked the desk
clerk, "Can you tell me where this meeting is, this fibrinolysis meeting?" She didn t
know what fibrinolysis was; she just gave me a quizzical look, and said, "Oh, the
meeting of the doctors!" 1 said, "Well, yes, that s close enough!" [laughs] She said,
"That started today," and my heart sank, because I thought, oh my God, 1 got the wrong
day, it s the time change, I m totally messed up. I ran up to my room I had a hot-pink
sweatshirt on, just similar to this, and black pants, and threw my suitcases down, didn t
do a thing, and ran over to where she said to go. I peek in the room, and there s a big,
conference-room table with about thirty guys 1 didn t see any women in there all
sitting around this big table. And I sat in the back, I just walked in the back and they
started looking back, and I took out a notebook and started taking notes. And they kept
looking back at me but continued with their talks. About ten minutes after 1 got there,
Desire Collen gave a talk and said that he had melanoma cells that make this substance
he called tissue plasminogen activator. He said he had an antibody against it, and he had
purified protein. And he had treated a woman who had deep-vein thrombosis, a blood
clot in her leg, and the blood clot dissolved with this substance. And here was exactly
what Genentech had sent me to hear.
Hughes: Right.
Pennica: And Genentech said, "See if anyone has a cell line. We need the cell line to extract the
RNA; we need purified protein, we can try to determine the structure of it by
conventional protein sequencing methods, and an antibody is also very helpful, because
then you can detect the protein in various ways."
Hughes: So Vehar had told you all this?
Pennica: To try to see what they have, what reagents; these are the things that we needed to clone
anything: You need a cell line, you need antibody, you need pure protein. So I was tuned
to those things, and here this guy got up there and said we have all these things, and I m
taking notes. Then there was a break, and somebody came and said, "Can I help you?"
And I say, "Oh, I m really sorry, my name s Diane Pennica, I m from Genentech, and I
am late for this meeting. I don t know how I messed it up." He said, "Oh, no, this is not
the real meeting, this is a pre-conference session." It turns out I didn t know this at
the beginning of the meeting they have all the hotshots, the head groups of the labs, or
the famous people to talk about their work, [laughter] The real meeting was three
hundred people; there was only about thirty of them in this little conference room. They
said, "But you re welcome to stay." I said, "Great!" He said, "And, you can join us for
dinner if you d like later!" [laughs] They had dinner at a castle, and I got to meet Desire,
face to face. They were all M.D.s, I m sitting at this table I ve been at Genentech one
month I m describing to them what cloning is on napkins, all excited, and saying,
"This is what we can do, and this is what we ve done." I had never cloned anything in
my life, but I didn t tell them that.
Hughes: [laughs]
Pennica: And I said to Desire, "Would you consider having us try to clone t-PA?" And he said,
"Oh, it s a huge molecule, the only thing you guys have done is growth hormone, that s
really tiny, we know how big t-PA is." And I said, "We can do it!" I had no idea if we
could do it, but I figured, you know, be enthusiastic, and that can t hurt. So we talked at
dinner, and I met the guy who first crystallized urokinase, a really famous man, I don t
even remember what his name is now. But I met all these guys who were instrumental in
the fibrinolysis field. The real meeting was three hundred people, and I might not have
ever met Desire, I might have been too shy to go up and speak with him, I might have
only heard his talk, gone back and reported, "There is a guy who has the reagents we
need to clone t-PA." But I
Hughes: Yes, but you short-circuited
Pennica: I short-circuited everything. I got face-to-face with this guy, and we talked, and we got
along. The real meeting was almost a blur because I was so excited that I had met this
guy. I don t remember the other talks. I remember him getting up again in the real
meeting and talking again. I drilled in my head what he was saying.
Hughes: What was the attraction as far as Collen was concerned? Why was he listening to this
young thing? Who wasn t part of the in-group either?
Pennica: Right, I was not part of the in-group. Who knows? I really don t know. But I think that
he was intrigued enough with the technology, that he was also a very enthusiastic
researcher. He had a spark, and he wanted to make a big splash, and he probably felt, "It
doesn t hurt; we can try." And whether he knew much about I think he knew about
Genentech, because he said, "What has been cloned before is too small, is smaller than
t-PA, and I m sure it d be impossible." I believe there were posters, I remember this
very distinctly, and I think this was at this meeting I m positive, because it couldn t
have happened after that I was walking around these posters, and so everybody knew
I was from Genentech, and a guy walked up to me and said, "Oh, so you re from
Genentech," and I said, "Yeah." He said, "You re interested in I see you ve been
talking to Desire cloning t-PA." He said, "If we thought that it could have been done,
we would have done it years ago." I ll never forget his words, and I just thought he
was a famous guy, I can t remember his name, but
15
Hughes: Was he an academic?
Pennica: He was an academic, yes. So he said, "If we thought that it was possible, we would have
done it years ago." [laughs] Really?! So, 1 said, "Well, can t hurt to try." So, certain
words, I ll just always remember.
Hughes: It doesn t sound to me as though the received word, when it s contrary to where you
want to get, slows you down much.
Pennica: Right, exactly.
Hughes: That s about the second or third episode that you ve described, where you were told,
"No," and you went on.
Pennica: Right, right. Exactly. You don t listen to people.
Hughes: Where did you get that? Where did that idea come from?
Pennica: Who knows? Sicilian, strong-willed background, I don t know I just [pause] you
know, 1 try to tell my kids my kids meaning the people who work for me, because I
don t have children I try to instill in them the same sort of enthusiasm, and that s how
I hire them, based on whether I see a spark in their eyes when I m talking to them,
because if 1 don t, I m not interested. And most of the guys say, "Oh, I can t wait to
work in your lab!" Great, okay, fine, you re hired, [laughs] So I instill that in them. I ve
had people at work say, "Oh, that s never going to work." You don t know until you try.
A lot of people leave science because it s very frustrating it is a painstaking,
frustrating experience. You have to have a lot of tolerance for failure. But that one
success is just unbelievable. 1 would get excited about little successes. That s what I
think Paul Cohen instilled in me, that every little experiment builds on the next, and you
learn something from it. That you can get excited about a little progress. For example, if
today 1 add ten nanomolar magnesium in an experiment and it doesn t work, then you
try twenty. And then you try thirty, until it does work, or you change something else. To
me, that was always exciting. Many times an experiment didn t work at all, but I didn t
give up. That s the way I approach all experiments.
But I think it was innate, I didn t say, "Oh, I ve got to convey to them that I m not going
to give up." It was just my positive attitude, I just said, "Oh, sure we can do it!" Never
having cloned anything in my life, I had no clue whether we could do it or not. So,
Desire decided to work with us, and when I got back I said, "Look, I met this guy, by
accident," and Herb Boyer said, "Nobody could have done it but you, Diane." People at
Genentech said, "If you were male, they probably would have kicked you out." They
didn t ask me to leave that pre-conference session, because, they told me later, they
thought I was one of the scientists daughters, waiting patiently in the back of the room
for her dad.
Hughes: Isn t that something.
Pennica: And that s why they didn t say, "Please leave, this is a private meeting." So I was able to
stay there long enough, and I didn t seem threatening to them. I think that was probably
16
the key. [laughs] You know, I wasn t a spy, I wasn t a threat, and I was able to sit there
long enough to hear a lot. I was just in the right place at the right time.
Hughes: Right time, that s for sure. Well, then what happened, because now you were on a much
different track, 1 mean, what did you do about the urokinase?
Pennica: Well, can we
Hughes: Yes. [tape interruption]
Pennica: So you asked about urokinase.
Hughes: Yes.
Pennica: And how did this put a spin on that? Well, the two projects were very similar, because
they were similar molecules, but the advantage of t-PA was that it supposedly bound
stronger to fibrin whereas urokinase was more non-specific. So if you have something
that specifically dissolves fibrin which makes up a blood clot, it s less likely to do
damage in your body somewhere else. Because your body normally is forming blood
clots and dissolving blood clots, and there s a balance. When it gets out of balance you
have problems. If you have too much dissolving, you have bleeding, and you can have
hemorrhaging; if you have too little dissolving of the blood clots, you will form a blood
clot. So you form micro-blood clots all the time. When you exercise, if you run or jog or
do vigorous excercise, you make more t-PA in your body, and it keeps your arteries
clear. That s been known, they ve measured t-PA levels in your blood, and the levels go
up. So, they thought it was better because urokinase was very non-specific, and it would
more likely do damage in your body, so there was an advantage there.
Well, they sent me to this meeting to find out about this molecule, but Bob Swanson
was very interested in making sure we honored the Griinenthal contract, and he didn t
want me to take time to work on t-PA exclusively. He said, "I don t want you spending
so much time on t-PA, or much time." And I said, "But I can learn something. If I do an
experiment, and I find it can be applied to urokinase, this is worthwhile." I didn t think
it would be an issue because it s very frustrating to work on just one project, because
you can hit roadblocks and never go anywhere. And 1 said, "I ll make sure they both
move forward." But he wasn t too happy about it. But I did it anyway; I worked on t-PA
because I had made a promise to Desire that we would do this. I didn t want to let him
down, and I thought that this could be something good because he had said that t-PA
dissolves blood clots, and there was potential. So, we got all the reagents from him to
try it. But I think Bob might have been happier if I had done the urokinase project,
shown I could get that to work, then done t-PA. But I did them side-by-side, and t-PA
went faster.
Hughes: How does Herb Heyneker fit into this? Because you re still in his lab, aren t you, at this
point?
Pennica: Yes. I m still in his lab. He was happy about it. I mean, you know, the harder I worked,
the better. He was another very enthusiastic guy, he always had a spark in his eye. I ve
been very lucky: There are so many people in science who have terrible Ph.D. advisors,
terrible graduate school experiences. I have been so fortunate because everybody I ve
17
been involved with has been excited about science. But there s some dull people in
science. I ve met a lot of them, (laughs) And I m saying, "Oh God, I m glad I don t
have to work for him!" Or her, or whatever. Herb was always saying, "This is great, this
is exciting!"
Hughes: Was he considering that the urokinase and the t-PA were yours? He wasn t directly
working on either project?
Pennica: No, he was more an administrator; he didn t work exclusively in the lab, but he was
working on growth hormone at the time, I think, growth hormone and insulin. So these
were my projects, and Bill Holmes and 1 were working on both of them at the time.
I lughes: How did the division of labor go there?
Pennica: This eventually split us up; 1 eventually left Herb s lab because Bill and I just couldn t
get along. It was just a conflict of a battle of wills, you know. Bill Holmes was doing
the cloning part of it, I was doing the RNA part, and I wanted things to go faster. He
was probably doing everything right, but I thought I could do it better I don t know
why, I just thought 1 could do it better. He got upset, and I got mad, and I said, "I want
to try this," and Herb says, "Oh, I just think it s funny you two guys are fighting!" Well,
to me it was very upsetting because 1 wanted harmony, and 1 had never been in a
situation like this before, and it got so bad that I didn t want to go into the lab any more.
It was so upsetting to me. So 1 went to see Dave and no, actually, before I went to see
Dave, Goeddel came to me 1 had had success with doing some aspects of the t-PA
project, and he was following it all along, and he said, "Would you try gamma
interferon? We re trying to clone gamma interferon." So this was a third project. I said,
"Sure, I ll help you out." And I thought, "Why not? I could do it on the weekends." I
was working seven days a week at that point. He had people in his lab working on the
RNA, but they probably didn t have the expertise that I did. There was a race with
people in Switzerland, I believe I can t remember the names, you probably know.
Hughes: Biogen?
Pennica: Biogen.
Hughes: Charles Weissmann?
Pennica: Weissmann, yes, Weissmann. There was a race, and [pause] Dave tried for about a year
to do this. They wanted to extract the RNA, inject the RNA into oocytes these are frog
eggs take the medium the eggs were sitting in, and put it on an assay and see if they
could measure interferon activity. They had been trying that for a long, long time,
couldn t get it to work. Dave asked me if I would try to extract the RNA, because he
knew that I had experience in it. So I used my famous gels and I separated RNA, like I
had done in my graduate school days, and 1 sliced the gel into different fractions, we
had them injected into the oocytes, we got activity, just like that, first try.
Hughes: Oh, really isn t that amazing. Had the RNA probably degraded in the previous
experiments?
IS
Pennica: Probably. Yeah, and I was so meticulous. 1 was as meticulous as can be, and everything
was fanatically done. That s why I didn t have time to go down the hall and find out
what everybody else was doing because 1 was so focused on what 1 was doing that it
worked the first time. Dave then took the RNA after we got activity. He worked non
stop. He took the RNA, he made the cDNA, and cloned it, and that s how we got to be
first.
Hughes: There s another intense person.
Pennica: Oh, God. [laughs] I worked for him for thirteen years! Yes, very intense.
Hughes: How did that work, I mean, two intense people in the same lab?
Pennica: I learned a lot from Dave, 1 learned a lot, so [laughs]
Hughes: So, he knew then that you could do something.
Pennica: Yes, he was all excited, so when it got to a head between Bill and I, because we were
arguing every day, and he was mad, and I was mad. I was so upset, I just went to Dave,
and I said, "You know, I can t work there anymore." He said, "You can come work for
me!" I took the t-PA project with me, which Herb was upset about, and I said, "You can
have urokinase because that s what you started on before I got there, and
Hughes: And it still wasn t absolutely clear which was going to turn out to be the better
Pennica: Exactly.
Hughes: So it wasn t that
Pennica: No.
Hughes: Well, I should ask you: Was Herb thinking, "Here goes that woman and she s taken the
better of the two projects?"
Pennica: I think he might have thought that; he might have, but I don t know what his thought
was at the time. He may remember differently, but this is the way it happened. I was so
upset, because I loved working in the lab, and I just thought, "This is so upsetting, 1
can t go in and fight with this guy every day." But Herb thought it was funny, and Dave
says, "Oh, you can work in my lab!" I had proven myself already to him. So I said, "No,
I m taking t-PA with me I got Desire, 1 made all the contacts," and I wanted to take
t-PA with me, and I stopped working on urokinase at that point. So, I went into Dave s
lab, and that s what I worked on a 100 percent for a while.
Hughes: And did it really move faster?
[Tape 2, Side A]
Pennica: Yes, but it did take a long time. No, I worked nonstop for two years, and basically didn t
have a life. I worked seven days a week, I worked from eight in the morning till,
sometimes, one in the morning, doing experiments. And then rumors kept coming that
19
other people were working on t-PA. Kabi, Cold Spring Harbor, Genetics Institute were
all trying to do the same thing, and that made me work just that much harder because it
was a struggle. We had trouble initially trying to identify a true clone, and we were
doing these experiments where you had to sequence some of the positive clones that
came up on these films, sequence DNA from the clones, and Bill Kohr 1 don t know if
you ve heard his name
Hughes: Yes, yes.
Pennica: He s one of the inventors on the t-PA patent. He is a brilliant man; he was the head
protein chemist on the project, protein sequencer, and without him we couldn t have
done this project. He is such a great guy, smart, smart guy, and no ego. He s just
brilliant. And he s told me stories that are just wonderful, too. His bosses would say,
"Try this, this, and this," and he would ignore them. He said, "If I did what they said, we
would not have had the t-PA sequence." So, he s got some great stories, too. He was
sequencing the protein, and he would bring me little stretches of the amino acid
sequence that we would make probes from. Now, when you sequence something, if the
protein is 100 percent pure, then every fragment of sequence that you get is going to be
from that protein; but if there s 1 percent contamination of one or more other proteins,
you may get a little stretch of protein sequence that s not t-PA. And so you could make
a probe to some other protein that we don t know the sequence of, and you re going to
be spinning your wheels for a long time, and not get what you want.
And so Bill kept giving me sequences, and I would sort of remember them. I would
memorize the sequences of some of these protein fragments. When we were looking for
the very first clone, Peter Seeburg was sequencing them at the time, and he would come
to my lab, put a piece of paper down, and say, "Nope, those clones aren t t-PA,"
because he would compare it to the amino acid sequence that Bill Kohr would give me.
The computer system was just getting up and running at Genentech, and 1 think he was
doing a lot by hand. Well, one day, he had sequenced a lot of clones that just didn t turn
out to be t-PA, and it s very frustrating, it s disappointing and upsetting because the
spots would be blazing on the piece of film, and you d think, "Oh, that s got to be it!"
Well, some of those were probably probed with the wrong thing, others may have just
been non-specific stickiness on the filter. Then, one day. Peter came and threw some
paper down on my desk and said, "Nope, we don t have it," and after he d left, I looked
down, and I had memorized one of the things 1 saw, the sequence W-E-Y-C-D, I think
that s what it was. I said, "Oh, my God, that s one of the sequences that Bill gave me!"
I went in to Dave s office, and my hands were shaking, and 1 said, "1 think we have it."
So that was the first clone, and he said, "Really?!" And I was shaking, I was so excited,
because we had gone through clone after clone. Peter missed it. Either the computer
system didn t find it, or he didn t remember, or we didn t have that sequence in the
database, I don t know what the reason was, but he missed it. And 1 just looked down,
and I saw that because I had remembered one of the sequences that Bill had given me,
and I was so excited.
Then things moved very quickly after that, and again, the best way is to look in my
notebooks to see the sequence of events because I can t remember. But Dave was great
because he knew everything about cloning, he told me what to do next because he was
the expert. He knew what things had to be done. All this was new to me, so it was a
learning experience for me. But he knew how to do it my hands did it, and he was just
20
telling me, "Try this, try this, this, this, this." And most of the time it worked, but a lot
of the time 1 did five things at once trying to get something to work. He s a smart man,
and he knew exactly what needed to be done, but it took a lot, a lot of work to get that
first clone. I worked solidly for another five months, seven days a week, to try to get the
final piece of it. The first clone we got was missing about a third of the molecule up at
the 5-prime end, and we tried so many different things that just did not work we
couldn t get it because we think that maybe the RNA is twisted in a little hairpin. It
might make it difficult for the enzymes to get through and make cDNA. We don t know;
we ll never know to this day why it was so tough, but we tried a number of things. We
ended up getting a genomic clone to probe a library, and again, I learned a lot about how
to clone in that stretch of time, but finally got the 5-prime end.
We started writing up a paper, and again, there was another disagreement where I was
mad at everybody, because Gordon Vehar went and told Bob Swanson, "I don t think
we should publish this paper." And here I had worked for two solid years doing this, and
I said, "What do you mean?" We had a policy to publish papers. He said, "Because the
competition will have the sequence, and they ll be able to catch up." I was devastated, 1
was so upset. I went to Bob and I said, "How could you do this?" Gordon was doing
assays for t-PA, but he wasn t as
Hughes: It wasn t his project.
Pennica: It wasn t his project. And I was so upset with him for the longest time. A few weeks
later, somebody called me and said that, "We have a paper from Kabi that is a partial
clone of t-PA" they were going to publish it in PNAS [Proceedings of the National
Academy of Sciences], and I was devastated. They said, "We want to know if your
paper, because we know you re working on it, have you cloned it yet, have you " We
had cloned it.
Hughes: This was the publisher, now?
Pennica: No, this was a reviewer he shouldn t have done this, he never should have called me. I
don t know who it was, but he said, "If your paper s already in press, I ll reject this
paper, I won t accept it." I said, "I have to be honest with you: We haven t submitted it
yet," because Genentech was holding it back; I didn t say Genentech was holding it
back. I was so upset, and then I called Bob, and I yelled at him. [laughs] I really yelled
at him. And he had Human Resources call me, and he said, "Try to calm Diane down."
They said, "Why didn t you call us first?" I said, "I don t have a problem with you; I
have a problem with Bob!" [laughs]
Hughes: Good for you, good for you.
Pennica: I yelled at him. I said, "Bob, you haven t been working as hard as 1 have, I ve been
working seven days a week I want to publish this stuff." I said, "And a journal editor
just called me and said another paper was going to be published we re going to get
scooped." I said, "I know they don t have a full-length clone, but we re going to get
scooped." And then they let me submit it to Nature, because another paper was coming
out.
Hughes: How long was the publication delay, do you think?
21
Pennica: I don t know, that I can t remember. 1 just remember calling up Bob and yelling at him.
No, 1 think I did it in person. 1 was shaking, 1 was so mad. (pause] He has probably
never had anybody yell at him like that [laughs] 1 said, "How could you do this 1 .
Because we don t want to get scooped." Well, [pause] 1 don t know, it was so
emotionally charged.
Hughes: Did this have anything to do with Swanson s original skepticism or if you could put it
that way, about why you should be even concerned with t-PA?
Pennica: I don t think so. I think at that point he knew, because he d heard about all the rumors of
everybody trying to clone t-PA, that it might be a big thing, and that if we published the
whole sequence, then anybody in the world could get it.
Hughes: So he was getting it, that this was a hot thing.
Pennica: Yeah, yes. So Gordon convinced him, "Oh, we shouldn t publish this." Oh, I was so
mad. I was so mad.
Hughes: And it was published in Science?
Pennica: Nature. Kabi did publish theirs, but it was a partial clone, so it wasn t the full-length
clone.
Hughes: So you got the credit that you deserved?
Pennica: Yes, yes. And the Kabi guys I actually met them they came to Genentech, they were
in town one day, and they said to me, "How did you do it? It almost killed us!" I said,
"It almost killed me, too." I said, "But I probably worked harder than you guys." I said,
"It was the toughest thing I ve ever worked on, and the fact that I heard, that there was
rumors that you guys were working on it too, made me work so much harder." And I
said, "I m not surprised you didn t get a full-length clone, because it took me forever to
get that final bit."
Hughes: So they d had the final problem with that final end.
Pennica: They had a problem as well. You know, I think we scared a lot of people because I think
they thought that we had an army of people working on it. I said, "No, it was just me
doing the cloning at that point." There were other people Bill Kohr was instrumental,
Gordon was doing assays, Dave was the director he was the one giving the
instructions.
Hughes: And then what happened? Did you have anything to do with the development, the scale-
up, any of that part?
Pennica: No, no. I did not. I expressed it in E. coli, I expressed it in mammalian cells, and then it
had a mind of its own. I mean, the whole project was taken over by other people.
Hughes: I see.
Pennica: And I moved on to the next thing.
22
Hughes: Now, what about the fact that Genentech had this mammalian cell expression system?
We re talking now very early eighties, right? Do other people have mammalian
expression systems, or is this giving Genentech an advantage, that you already have this
system in place? Or was it in place for t-PA?
Pennica: I think they must have done some mammalian cell expression, but I don t know, I really
can t they used CHO cells and 293 cells-
Hughes: I think interferon.
Pennica: Interferon probably was, but I really don t know, at the time. One thing that I forgot to
mention, that was another high point, is after the cloning, I submitted an abstract to a
meeting to give the talk on the cloning of t-PA, in Switzerland? I can t remember where
it was. It was going to be the announcement. I had to write an abstract that was very
vague we re trying to do this, this is how we re trying to do it. I couldn t say anything,
and our lawyers said, "No, you can t say it because it would be like a public
announcement."
Hughes: Oh, absolutely. So the patent hadn t been filed yet?
Pennica: The patent hadn t been filed? No, the patent had been filed, but not released, so it s not
public knowledge that we had done this. So, I had to make a very vague abstract. The
meeting accepted it but they put me as the last speaker on the last day; there was five
days of talks. I was the talk right before noon, and then there was a clinical session,
which was not science stuff. And I said, "Why d you put me on last?" I said, "I hate
being last." One of the guys I had met at that first meeting in Malmo said, "Because we
don t know what you re going to talk about, and you didn t give any information." I
said, "Well, 1 can t give any information I guess you ll see when I get up there and
talk whether it s of interest." I was so excited because I had cloned t-PA, and Desire
was there at the meeting, and he knew that we had it. I ll bet that he told people, but 1
don t know for sure. I got up, and I gave my talk, and they set timers when you give a
talk, usually at meetings, because they don t want people going over. Halfway through
my talk, I had just flashed up the structure of t-PA, with all the kringles and all the dots,
I had put on a board, the timer goes off. I looked at the audience, I said, "Should I stop?"
The whole audience says, "No!" [laughs] Everybody started to laugh, and I continued.
At the end of my talk, they gave me a standing ovation, which is unheard of at any
scientific meeting. I couldn t believe it. Somebody came up and said, "Are you one of
the marketing people? You gave such a great talk." I said, "No, I m the scientist who
did the work!" They said, "But you were so polished." I said, "Because I practiced for a
month to give it." But I practice and practice, because I m so nervous, that they thought
a non-science person gave this talk.
Hughes: Isn t that amazing?
Pennica: But that was really a high point.
Hughes: So people did come, probably because Desire had said, "You d better get there."
Pennica: Probably, but the room was probably three hundred people, and one guy had put up a
I think from Genetics Institute, which made me so angry he put up a camera in the
23
aisle, and started filming it. One of the guys it might have been l-lliot (irossbard
who went up to him, and said, "What are you doing?" And he said, "I m filming." He
took pictures of every one of my slides, which was pretty (laughs] lousy.
Hughes: Yeah, I m surprised that s even allowed.
Pennies: Well, they stopped that after, but he took pictures of all my slides. Then somebody
came up to me and said, "Well, how are you expressing it? Do you have expression in
yeast, do you have expression in E. coli, do you have expression in mammalian cells?"
And I felt like I was being pumped for information.
So right when 1 gave my talk, Genentech issued a press release that we had cloned t-PA,
at the exact same time. That was pretty exciting.
Hughes: 1 can imagine.
Pennica: It was too bad that my talk was on the last day because I would have liked to talk to
other people at the meeting. Because, if you re first, people come up to you and talk to
you throughout the rest of the week because they want to know more about what you ve
presented. The organizers just said, "Well, we didn t know you had anything important
to say."
Hughes: What is Desire Collen like as a personality?
Pennica: Oh, he s great; he s a teddy bear! [laughs] He was always sweet; every time he came
here, he d bring me a little box of Belgian chocolates. He was excited, too, about the
work he got the equivalent of the Nobel Prize in Belgium for the t-PA work.
Hughes: Is that so?
Pennica: Yeah. I don t know what the name of the prize is, but it s the equivalent. He now has a
whole institute, 1 think, that he runs, and he s become very famous over there. So, he s
great.
Hughes: Do you know the history of how he became interested in t-PA?
Pennica: No, I don t. That s a good question. Was it fibrinolysis? Maybe he had been studying
fibrinolysis his whole life, and he isolated a substance because he wanted to try to
dissolve blood clots. 1 don t know how he became interested. They didn t know for a
long, long time what in your body dissolved blood clots, or even if there was such a
thing. I have a series of slides, that, if we are going to get together to talk again, that I
can show you, that details the history of this, which is so exciting you actually read it,
you read part of it in that book chapter about how they discovered that it was actually
worth something.
Hughes: I know Marsa gives credit to Peter Rentrop?
Pennica: Yes, right.
Hughes: Is that the way you pronounce his name?
24
Pennica: Yes. But I did a talk detailing the history of it but whether Desire he was probably
looking for things that dissolved blood clots because he found that this melanoma cell
line produced a substance that would dissolve blood clots and then subsequently tested
it on a woman who had a deep-vein thrombosis.
Hughes: And as so often is the case, 1 saw some parallels with the interferon story, that people
have known about these substances for decades, but because there is so little of it.
Pennica: Right.
Hughes: It holds the science back.
Pennica: That s right. If you can t get pure protein, you can t clone it. Well, you can now because
if you have an activity, you can. They re called expression-cloning projects, but back
then, our techniques were so archaic that I can t even imagine trying to do some of the
things that we did back then. It takes a week now to do things that took a year back then.
Hughes: As we know again from the Marsa book, there was a lot of controversy, particularly in
clinical trial and FDA approval stages. Were you watching that? Here was your
substance, moving slowly and with considerable setbacks toward actual clinical use.
Pennica: I watched it briefly, but again, 1 was so entrenched in my next project, which was TNF
[tumor necrosis factor], that I was almost oblivious to what was happening. I just
thought, "Well, 1 hope this works," and I kept hearing things that were going on but
didn t follow it that closely because I was, again, working on stuff on the next project.
That was just my intensity; 1 can t think of anything else, [laughs]
Hughes: Shall we save TNF for next time?
Pennica: Sure, yes.
Hughes: Let me just do you have a few more minutes?
Pennica: Oh, yeah, of course.
Hughes: I do feel a little uneasy, since I don t ask the men this
Pennica: Oh no! [laughs]
Hughes: But 1 think it s relevant to ask you how it was to be a young woman in what seems to me
to have been not only a really predominantly male organization, but also, in the early
days, somebody has even described it to me as kind of like a fraternity house, you
know.
Pennica: Yes.
Hughes: [laughs] So how was that?
Pennica: Good and bad, in a way, because I didn t feel part of a group, the gang. 1 just didn t. But
in a way it didn t matter, because it made me focus more, and I [pause] you know, I
25
think anyone can succeed despite the tact of being a woman. 1 think it s tough because
there are a lot of things that have happened that 1 feel are not as fair as they could have
been, but I have been successful and done a lot of things because 1 keep my nose to the
grindstone, and 1 try not to get upset about a lot of things. Luckily 1 stayed when it got
so uncomfortable in the lab, when Bill and 1 kept disagreeing Bill Holmes but I m
glad 1 stayed because I don t know what 1 would have done. I would have gone to
another company, 1 guess, but it was just too much fun here.
Hughes: Do you feel that because you were a woman you had an extra burden of proving
yourself?
Pennica: I think so. I think that s very true. But I also think I got into that meeting because I am a
woman.
Hughes: Yes! [laughs]
Pennica: They said, "If you were a guy, we would have kicked you out, because we would have
thought you didn t belong here."
Hughes: It cuts both ways, doesn t it?
Pennica: Right, yes. It s been tough at times, but it s
Hughes: Do you see any difference in well, this is probably an impossible question do you
see any difference in the way you think about and actually do science because you are a
woman?
Pennica: Not at all.
Hughes: No.
Pennica: No. I don t think that makes a difference at all because my style is probably different
than any other style. I m more fanatic than most about doing everything perfectly and
that probably increases your chances of success. A lot of people say, "Oh yeah, I tried it
this way today, the next day, tomorrow, I ll do it a different way." But if you don t do
an experiment the same way day after day, and you know what works, don t change
anything. This is what I drilled into my people, but I don t think it has anything to do
with being male or female. It s just your personality. I always loved the lab work, I
thought it was so much fun, I always got excited about it. But that s not a male/female
thing because 1 think there are men who a colleague of mine, Michael Marietta, who
is now at UC Berkeley, he won the MacArthur Genius Award he went to Fredonia
State. He was in the same classes with Dr. Fox, and all the professors I had, and he just
recently this year got in touch with me. I hadn t seen him in twenty years.
Hughes: How funny.
Pennica: And he won this award. He has a spark, he is another guy that works intensely at
science. You just can t be set back, because there are a lot of problems but male/
female? No, 1 don t think that s I think that was an easy question!
26
Hughes: [laughs] Do you still do hands-on science?
Pennica: Not any more. I have two people working for me and it s hard because I think I want to
have a life now, and 1 didn t before, because science is so demanding. It s very tough
mentoring people and working in the lab at the same time. Because when I m in the lab,
I don t want to talk to anybody; I don t even want to be interrupted for a second because
you can be distracted too easily. If you re only supposed to spin something for thirty
seconds, you can ruin an experiment if somebody comes and talks to you and you spin it
longer. So I was probably thought of as antisocial, because I didn t walk around,
because I paid attention so carefully. I resisted for a long time to get somebody to work
for me. Dave would say, "You ve got to get somebody to work for you." I don t want
anybody, I don t want anybody. I thought it was too tough to teach somebody and work
at the same time. And it is. I usually got kids right out of college, who wanted to go to
medical school, wanted to go on to graduate work, because I knew they would be
ambitious. But then I had to start teaching them, "Here s how you pull a filter, hold a
pipette this way." I thought, "How can I do work in the lab, because they would be
asking me questions every ten minutes." And so I slowly, slowly phased out of the lab
and now I don t work in there anymore. But I miss it.
Hughes: I bet you do.
Pennica: It s just different.
Hughes: When did you marry, and is that one of the reasons that you began to cut back?
Pennica: No, I am now divorced, and I m very, very happy. I m very happy with my life right
now. I got married in 1 990.
Hughes: So after all the t-PA.
Pennica: Right, after all that, because I was too busy before. It was just too busy. Yeah, I don t
know if I could there s a lot of women who can do it all, have kids, have a full-time
career, have a good marriage, and I didn t. I was 100 percent focused on work. Now I
really want to have a life. I still love the science, and get excited about it, but I can t
work here a hundred hours a week like I used to. [laughter] It was too much.
Hughes: That seems reasonable!
Pennica: Right. My mother died very early, she was fifty-nine, she died of colon cancer, and
[pause] life s too short not to have fun, and I just hadn t had fun for a long time. I had
fun in science, I will never regret that, but one thing I didn t tell you about, but this
was in the book. This is a Christmas card that I got from Mike Blum no, that s not in
that book. That s different, that was about the first heart-attack patient I met. Then I got
Christmas cards and other letters. I have things tacked up all over my wall from
patients. That s only a handful of the things that I ve gotten.
Hughes: And what is the Mike Blum?
Pennica: He had a heart attack, and he got treated with t-PA, and I just got this random card from
him. The card says: "Dear Dr. Pennica, Thanks for helping to save my life. Mike
27
Blum." Boy, when I have a bad day, or 1 think I m not being treated well, I look at that,
and say. "Well, there s no other guys who have thai tacked up on their wall."
Hughes: Well, let s end with that story of meeting the patient. Is it really true that it was the day
of the party, because we didn t talk about the party, either, the celebration ?
Pennica: No, it wasn t the day of the party, 1 don t think. I don t know why he came here, I think
he wanted to see the company that made t-PA. Steve Birnbaum came to Genentech, and
I was in the hall, walking very fast like 1 always do, trying to get to the lab, or
someplace. One of the marketing guys said, "Oh, Diane, I want you to meet somebody."
1 said, "Sure." Again, 1 was always in a hurry to get to the lab. He said, "This is Steve
Bimbaum. He is the first heart-attack patient to be treated with t-PA." I said, "Oh my
God!" Steve didn t know who 1 was, and he said, "This is Diane Pennica. She s the
woman who cloned t-PA." And Steve immediately just grabbed me and hugged me, and
said, "Thank you! Your drug saved my life." Unbelievable.
Hughes: What an emotional
Pennica: Oh, it was great, absolutely great. That one experience was worth any aggravation that
I ve suffered [laughs] over the years.
Hughes: You ve spoken a lot about your passion for the science. Was this kind of icing on the
cake, the fact that, yes, the science is really exciting, but look what it does for people?
Pennica: Right, and I never thought that I would ever meet somebody clinicians, M.D.s meet
patients all the time, they re always getting thank yous, and things like that. As a
researcher, you never think that you re going to meet a patient, ever. 1 feel very
fortunate because most people who work in science are never going to get that
opportunity. They re going to work on things that may never translate into something
that hits the clinic, and to have something like that happen is incredible. Somebody
came into my office last week and said, "I want to tell you that my uncle had a stroke,
and they gave him t-PA, and he is alive and well, and he wanted me to come thank
you." And I thought, "Oh my God!" So still now
Hughes: It s still happening.
Pennica: When t-PA was announced, my small hometown, Fredonia, had all these honors for me.
I got the Service to Humanity Award, I got asked to be a commencement speaker there,
I got lots of accolades. Then people from Fredonia would send me letters saying their
mother had a heart attack or "I had a stroke," whatever. So I ve gotten stuff like that,
too. It s really very rewarding.
Hughes: When you were doing the science so intensely, were you thinking in the back of your
mind about clinical application?
Pennica: Not really. You know, I sort of knew what it could be useful for, but never thought it
would ever have an impact like this because 1 didn t know our capabilities at the time. I
didn t know we had the facilities to actually take something all the way to clinical
practice.
28
Clearly, this is an amazing company, because it s done so much already, and I m very
honored to be a part of it. I am very fortunate that things have happened that who
knows why they ve happened. Because I m basically an unbelievably shy person, 1 hate
giving talks; I really hate it. And that s the only way you re going to get well known.
Well, I gave a lot of t-PA talks, but it s a struggle for me every time. To have something
like this happen, you know, "We want you to give a commencement speech" oh my
God! Two thousand people! [laughs] But I go and do it. 1 got asked to give a talk at
Vanderbilt University to five hundred graduate students and postdocs, and it was only a
fifteen-minute thing. I was terrified. This was just last year, and yet I did it.
[End of session]
29
INTERVIEW 2: JULY 23, 2003
(Tape 3, Side A]
Hughes: Today we re going to start with tumor necrosis factor (TNFJ, which I m going to leave
to you pretty much to tell the story, since 1 don t know much about it.
Pennica: So, tumor necrosis factor, which we call TNF, was first described in 1 975 by a scientist
named Carswell, as an activity in the serum of mice that, if you put it in vitro it would
kill cells, or in vivo, it would shrink tumors. We decided that that was something
worthwhile pursuing because it could be a cancer therapeutic. So, in 1983, 1 believe,
again I ll have to check that date in my notebooks, we decided to tackle this as a project.
We found a cell line, much as we did with t-PA, that produced TNF. You have to find a
cell line or some tissue that makes a lot of the protein that you re interested in, so you
can isolate the protein, determine the amino acid sequence by conventional methods and
make a probe to a portion of that sequence. You could then probe a library to see if one
of those clones has a piece of the DNA that encodes TNF. So that s the strategy we used
for t-PA, and we used a similar strategy for TNF.
We first looked for a cell line. We took a number of different cell populations and
assayed them for TNF activity. In a sense, we looked for cell killing, and we found that
a cell line called HL-60, a promyelocytic cell, produced activity. So we used those cells,
isolated some protein by conventional methods and got a preliminary protein sequence.
From that protein sequence we made a probe, we made a library, and we found some
clones. We got a sequence that, when we expressed it in E. coli, it did kill cells in vitro.
We injected mice with cells that made tumors, and found this protein was able to shrink
the tumors in these mice, which was very exciting. Unfortunately, this never went on to
become a drug because it had such potent activity and side effects, so we didn t pursue
this any further at that point.
Hughes: Did it get as far as human clinical trials?
Pennica: You know, I think it did, but again, I am so out of that loop. That s something that I
could check. I bet it did.
Hughes: Now, who s the "we" involved in this work?
Pennica: Again, the same people who were involved in the major players in the t-PA story: Bill
Kohr, who was instrumental in determining the protein sequence of the isolated protein;
and Dave Goeddel, again, with his wisdom, advising us on the best cloning strategy;
Glenn Nedwin was involved; Joel Hayflick, Peter Seeburg, Rick Derynck, Michael
Palladino, and Bart Aggarwal, they all made contributions. But I think the core was,
again, the same people who did work on the t-PA project.
Hughes: Now, are those cell lines you mentioned already in Genentech s possession?
Pennica: 1 think they were. We got these from the American Type Culture Collection, and we just
screened a number of them, including peripheral blood leukocytes, for activity. We
fractionated PBLs because they contain a number of different cell types. So, we were
successful, and we were the first, once again, to publish the cloning of TNF, which was
great.
30
Hughes: Who were your competitors?
Pennica: 1 think there were competitors in Europe. Charles Weissmann I think was trying to do
the same thing. Dave was always anxious to beat him. I think we beat Charles
Weissmann on the interferon cloning as well. I m not sure he was working on TNF, but
there was another group who published the cloning of TNF many, many months later in
Science. I couldn t understand this, because the journal should have said, "Well, this has
already been done," but they published it anyway. I can t remember the name of that
group, but I was upset, because I went to Dave and said, "How could the journal
Science let this be published? We already did it; they published the same sequence.
They just went about it using a different strategy." We did have competition, but we
were first. So that was great, very exciting.
Hughes: Who is responsible or how does it usually happen that a project like this gets off the
ground? Is somebody reading the literature and comes upon the work that you cited in
the case of tumor necrosis factor and said, "Well, maybe this is something that
Genentech should be developing?"
Pennica: Yes. At that time, they listened to anybody who had a great idea. I believe it was Rick
Derynck who said, "This is something that we should pursue: There s this activity. They
discovered that this stuff was in the serum of mice; it s probably something we can
clone." If it s not a single protein that s killing these tumors but a combination of many
proteins, then it s going to be impossible to clone. But I think Rick Derynck was the one
who came up with the idea, although he wasn t involved in the cloning aspect of it. So
he gets credit for that idea, and at that point we just decided to work on it.
Hughes: Did Swanson, or some of the other executives, have to give it an okay?
Pennica: 1 believe so; 1 think
Hughes: The scientists just couldn t take off, and begin a project.
Pennica: Right. But the fact that it was cancer-related, I think that was enough to warrant it being
pursued, and Dave Goeddel is very convincing, and he s usually right, 99 percent of the
time- probably a hundred, he ll say! This was something worth doing, and in hindsight,
it certainly was.
Hughes: Tell me about that, tell me how it became useful.
Pennica: Again, I don t know how far it got in clinical trials, but that s documented somewhere.
But right now, what s important about this is: Without the cloning of TNF, the drug
Enbrel would never have been developed that s for arthritis because Enbrel uses the
receptor for TNF to block TNF activity. The receptor was subsequently cloned also at
Genentech, as the target for inhibition by this drug. Enbrel could not have been
developed if TNF had not been cloned. That s pretty exciting. I get satisfaction knowing
that I was at least part of it.
Hughes: Were you involved in any way with the cloning of the receptor?
Hughes:
Pennica:
Hughes:
Pennica:
Hughes:
Yes, I was, and what s upsetting to me is that I was going to put my name on the
paper Dave put my name on the publication because I did probably two years worth of
work on it. It was very, very difficult, but in the end, none of the data that 1 did, even
though 1 worked for two years, went into the paper, so 1 felt that 1 should take my name
off. I m very upset that I did that now because people get on publications for doing
nothing. And that s upsetting. Dave said, "Oh, you shouldn t worry about what anybody
else thinks," and I was worried that they would say, "Well, none of Diane s data s in the
paper." He said, "But you worked, and your data helped to get this far." I could kick
myself, [laughs] But it s just a publication but 1 should have been on that paper and 1
took my name off. I should ve because 1 have notebooks filled with data on trying to
clone the receptor. It s not that I didn t work very hard on it; 1 just felt guilty because
none of my figures or anything else went into the paper.
I m assuming that in the progression of a project that areas that prove not to be
productive still contribute to the project.
Absolutely.
Because you know that then, this is not the way to go.
I was naive, I was naive in thinking that I was worried about what all the other authors
would think. Silly me. [laughs]
Is there usually a lot of discussion around who should be authors and how they should
be ranked?
Pennica: Yes, a lot of discussion, and it gives people a lot of anguish and heartache for people
they want to put on that they don t, or vice versa. Other people who think they ve
contributed a lot sometimes don t get on. It s a tough call. My strategy has usually been:
Put everybody on the paper who has helped. Because it makes them feel terrific, they ll
want to help you again. As a result, I ve really been fortunate because a lot of people at
Genentech say, "Oh, I m going to help Diane because she put me on this paper and this
paper and this paper," and they usually do make a substantial contribution, so I have no
problem. But in the beginning, there were some papers that I thought, "Why is this
person included on this publication?" because he or she did not do enough to warrant it.
That s why I felt that I should take my name off the TNF receptor paper because none of
my data went in, even though I worked maybe harder than some of the authors, but
hindsight s 20/20! [laughs]
Hughes: Yes, yes, it definitely is.
Pennica: And Dave convinced me, he said, "Oh, don t worry about what anybody else thinks."
He said, "You should keep on it," but I said, "No, I don t want to be."
Hughes: Does Genentech in general follow what I believe to be the norm in that the last author is
usually the lab in which the work was primarily done, and the first author is the person
who did the most hands-on work?
Pennica: Yes, right.
32
Hughes: So those two are probably not terribly difficult to decide, it s just the authors in between
and how they should be listed.
Pennica: In general, yes, that s true. Yes, because it was always nice to see "Pennica, et al."
Because when you reference, it s the first author, usually. Or if somebody in a journal is
referencing the paper, they may say, "GoeddeFs lab," because he would be the senior
author. But in this case, different from t-PA, Dave went on to give all the talks on TNF.
He announced it in a conference. 1 did the t-PA stuff; he announced the TNF work even
though I did the majority of the work here, [pause]
So anyway, it was another exciting thing because we were first once again, and that was
the important thing and that was very important to Dave, and it was important to me,
too. 1 liked being first; 1 liked to be the first author on a paper because I ve been beaten
before. I ve worked on things that I ve been scooped on. You open up a journal, and
you ve worked for a year, a year and a half on a project and find out they ve cloned
exactly what you re working on. It s so depressing because you could publish it months
later, like they did for TNF, but it doesn t have the same impact.
Hughes: Do you think that being first was a stronger drive at Genentech than most other
organizations?
Pennica: Well, I think, for patent reasons, it was important to be first. I never thought of it that
way because I didn t know at the time how patents protect you. But scientifically you
wanted to be first, because, again, nobody remembers number two! [laughs]
Hughes: Exactly.
Pennica: I ve gone to meetings, and they would say, "You re the woman who cloned t-PA, you re
the woman who cloned TNF, you re the woman who cloned p53." It s a great feeling.
Hughes: I bet. Well, tell me a little bit about the patent process from a scientist s standpoint. I
mean, when you do do something, and you know you ve succeeded, what happens then
in terms of the intellectual property protection?
Pennica: Well, the lawyers are contacted, and now
Hughes: Would you do that if you re the lead scientist on the project?
Pennica: Yes. This all happened in the background, usually. For TNF and t-PA, the lawyers were
busy working on it, and I was busy working in the lab so I just conveyed what data I had
to them, but I was not involved in writing the patent, or anything else. So I considered it
more of an annoyance, because they were taking up time away from the lab.
Hughes: Well, maybe let s use TNF, then, as sort of the model because there you must have been
directly involved in writing up the patents, weren t you?
Pennica: No, I wasn t, and I don t think I m even on the TNF patent.
Hughes: Not TNF, I meant t-PA. Describe what that entailed.
33
Pcnnica: The lawyers would come to you and ask you for the sequence; they would ask you for
the protocols, the methods, exactly how you did it, what cell lines you d used, what
antibodies, step by step.
Hughes: Now, was this Tom Kiley?
Pennica: Tom Kiley, yes. He was nice to work with; he was great. 1 think he was excited about all
this, too.
Hughes: Do you discuss kind of a legal strategy? 1 mean, how to present this, or is that in the
bailiwick of the attorneys themselves?
Pennica: The attorneys do that.
Hughes: So you re just giving the scientific data.
Pennica: Right, and 1 think they were learning as they went along because this was new territory.
I don t know how many molecules had been patented that had been cloned before.
Somebody may have told you that in some of the interviews, what the status of that was,
but I don t know how many previously had been cloned, and you have to think of: What
if you can find another use for it? Can somebody else get a patent because they claim
that this molecule can be useful for strokes, and we don t put that in our patents? That s
something that Genentech had to leam, or maybe they were very savvy about it, 1 don t
know, it was all in the background. They just said, "Diane, give me this data, give me
that data." And I did.
Hughes: So it sounds as though you appeared at the right moment, but then very quickly went
back to your lab. [laughs]
Pennica: Yes, yes. I think 1 told you this in the hallway, maybe I didn t; 1 don t know if 1 told you
this. Maybe 1 was telling my RA [research assistant], 1 can t remember. But, at one
point, when 1 was working on t-PA, lawyers from London came did I tell you this
story?
Hughes: Not on tape.
Pennica: I was working in the lab. Tom Kiley hadn t called me in advance, and he said, "Diane,
we have the lawyers from London here, we need you to talk to them." And I said, "I
can t, I m extracting RNA." He said, "What do you mean, you can t?" I said, "I cannot
leave the lab, there s no way I m leaving." Because you have to start an experiment four
days in advance to get the cells ready, I wasn t going to leave. They couldn t believe it,
that I told them no. He said, "They re only going to be here for a day." I said, "I don t
care. I m not going to ruin my experiment." So I ll never forget that; 1 was just telling
them my experiments were more important than the lawyers from London. It s funny
now, but I was annoyed at the time. I was very upset because I was torn, but I wasn t
going to let my experiment mess up.
Hughes: Four days down the drain.
34
Pennica: Right. No, it s too hard. "What do you mean, you can t come?" "I can t come." Here s
this little woman telling the lawyers that she can t show up. But they never gave me any
advance notice that they wanted me to talk to them. If they had, I could have planned
my experiments differently.
Hughes: Well, it probably came as quite a surprise to them. Well, if that s enough on TNF, I
believe what was also going on at the same time was, you were working on p53.
Pennica: Yes.
Hughes: Do you want to tell a little bit about that molecule?
Pennica: Yes. I d like to read a little bit from a review that came out when this molecule was
nominated in 1993 as "Molecule of the Year." Because when we were approached by
Arnie Levine to clone this, back in 1983. The paper that we published was in 1984, and
Art [Levinson] knew Arnie Levine; Art did a postdoc in Amie Levine s lab, and Arnie
felt that we were the best at cloning things, so he came to us. Dave said, "Do you want
to do a side project?" And I said, "Sure." So there s a molecule called p53. 1 said,
"What s that?" But, I ll read a little bit from this.
Hughes: That s fine.
Pennica: Because it s an interesting story. It says, "Back in 1979, when p53 was discovered,
nobody would have nominated it for Molecule of the Year or even of the month. First
identified in association with tumor-causing viruses that don t cause cancer in humans,
this fifty-three kilodalton protein was simply one molecule among many, a discovery
that might or might not be important in cancer. Now, in a classic parable of how basic
research can have a profound impact on human disease, this dark horse molecule is
blazing a bright trail in cancer research. Found in both inherited and spontaneous
cancers, p53 is to date the most commonly mutated gene in human tumors and is one of
the star members of the tumor-suppressor gene family. Of the six and a half million
people diagnosed with cancer every year world wide, half have p53 mutations in their
tumors." Which I think is amazing, and nobody knew that at the time when we started
working on this project, back in 1983. "Sometimes called the guardian of the genome,
it s a leader in the body s anti-tumor army, helping to coordinate a complex system of
responses to the DNA damage that might otherwise lead to cancer." So if you get hit by
sunlight, for instance, or radiation, or tobacco smoke, your DNA will get mutated, and
p53 s function seems to be to suppress the tumor, the mutation effect of what these
damaging agents might do. "Like an emergency brake, wild-type p53 can halt cell
growth, and sometimes send a cell into a programmed spiral of death." Meaning, if the
cell s too badly damaged, instead of having that cell grow uncontrollably, p53 will kill
it, so it can t go on to form a tumor, which I think is amazing. So normally, it would act
as a tumor suppressor, but if the cell is too far gone, it says, "Kill this cell." "Ironically,
for the first ten years after it was discovered, it languished in the backwaters of research;
it was thought to act only as an oncogene, a gene that actively promotes tumor growth
because scientists were inadvertently working with a mutant form. In 1989, p53 s
fortunes changed; researchers found a point mutation in p53, and mutant p53 promotes
abnormal cell growth, the wild-type gene suppresses tumors. Suddenly, p53 became a
hot ticket in cancer research."
>5
When we cloned it in 1984, Arnie told me later, this was the first wild-type p53 gene
that was known to be published. There was another group, Moshe Oren, who published
p53 cloning, but 1 believe he published the mutant form, we published the wild-type. It
was only a few months before, or it might have been a year before, 1 can t remember.
So just to tell you a little bit more, more than fifty-one types of human tumors carry p53
mutations, 70 percent of colon cancers, 50 percent of lung cancers, 40 percent of breast
cancers carry p53 mutations. That s why it s become such an amazing molecule to
study. At the time that we were cloning this, I had no idea that this would be such an
important protein.
Hughes: That s interesting, isn t it? Because you re literally down at the test tube level, to use a
metaphor, as is of course appropriate; this must happen in many cases when there s not
too much known about this molecule, and then this whole different world opens up
during the clinical trials, or maybe before, 1 don t know exactly.
Pennica: This was before, just because they re studying tumors, and they re trying to figure out
what s going on. That s why when I have people working for me in the lab, 1 try to point
out the fact that even though you re working on something today, ten years down the
road this could be another p53 you don t have a clue because people don t know
enough about it. That s one of the reasons that I send out clones to everybody who
requests them, all around the world, because when we stop a project, even if it s not
going any further at Genentech, somebody else may discover something about this
molecule that we don t have the capability of doing, or we are focused on something
else. It s left my lab, so I figure, send it out to the whole world, they may find something
that s exciting. A lot of people don t have that attitude; they say, "1 want to keep
everything to myself, it s too much trouble to send out a clone." Well, it is, but I have
collaborators all over the world, and they have published a lot of very exciting work,
that may be important someday.
Hughes: And does Genentech have no compunctions about your releasing clones, for example?
Pennica: As long as we have patents on them, and we have a good MTA in place, Material
Transfer Agreement form, where the university, or whatever group we are working
with, signs all the papers that we need to have them sign, then it s not a problem. So I
can t send anything out. In the early days we could. Now they ve learned that we have
to protect ourselves and protect the universities. But I love it when people ask for the
clone, because I think, "Great, I m glad somebody s interested." They think it s
important; they think it s interesting. And they re working on some obscure thing that
could potentially lead to another therapeutic, which is great. We re not pursuing that
angle, so we might as well let someone else study it.
Hughes: Talk a little bit about the actual science, as you were pursuing it with p53. Was it
difficult to clone?
Pennica: No, I believe it was relatively easy, [tape interruption]
Hughes: Diane, you were saying off tape that it looks as though the protocol was pretty similar to
what you d use for t-PA and tumor necrosis factor, and you can describe that in a
minute, but I m wondering if, at this time, and now it s about 1984?
36
Pennica: Probably 1983.
Hughes: In 1983, okay. Is Genentech beginning to lose the advantage it had in being such a whiz
at the whole cloning this and the expression business? Are other companies and
groups and universities beginning to catch up?
Pennica: We had competition, clearly, because Genetics Institute, Kabi, Cold Spring Harbor,
they were all in the t-PA race to begin with, so they were trying to find the molecules to
clone. It was a question of deciding what was the best thing to clone. Now, maybe p53,
because it seemed so obscure, wasn t something that was in hot pursuit by anybody. I
don t know. But what was known at the time was that Moshe Oren and Amie Levine
had published the 3-prime untranslated region of p53 messenger RNA in 1983, but they
couldn t get the full-length clone. Amie asked Genentech for help in getting the rest of
the clone. So we took a cell line that they knew contained p53 messenger RNA. It was
the F9 embryonal carcinoma cells, and we had tryptic peptides that were specific for
p53, that we knew the amino acid sequence of, and we probed a library made from these
F9 cells. So Arnie had tried. He got a little stretch of the 3-prime end, couldn t get the
rest of it, so he came to us.
Hughes: And he was in academia?
Pennica: Yes. He was at Stony Brook at the time. Yes, State University of New York at Stony
Brook.
Hughes: Yes, I was wondering how much of a difference at this time, 1983, it makes to be in a
company such as Genentech, I mean, a biotech company that s a big point as
opposed to an academic laboratory, because you have people here who are specialized
in doing specific things. You are usually involved with the cloning.
Pennica: Yes.
Hughes: But then you have somebody else do the amino acid sequencing, et cetera, et cetera.
Pennica: Right. We have divisions of labor.
Hughes: Right. Divisions of labor. But I would think that you d have to be a pretty big academic
lab in order to have all that expertise in one place.
Pennica: Probably, yes. Although people kept coming to me later at meetings, saying, "We could
never compete with Genentech. It s such a powerhouse. You must have a hundred
people working on a project." And 1 said, "Oh, no. Three, four, maybe." They don t
realize that we had the best in the world Dave Goeddel, who knew how to clone and
passed on his knowledge to me, and so I got the best teacher. And Bill Kohr, who
people outside Genentech had said, "He s the best protein sequencer in the world." So
with that in hand, you don t need that many people. You just need to work hard. And I
think these outside people were intimidated, thinking, we can t work on this because
Genentech s working on it. That s what I got from a lot of academic people, and I said,
"Why not?"
37
(Tape 3, Side B]
Pcnnica: People would assume they never could, compete with us because we had hundreds of
people working on a project, but that wasn t the case. We just had a small group who
worked like a hundred people.
Hughes: Yes, yes. And who also, as you were pointing out, were in most cases the best in the
field.
Pennica: Yes. I think we had the best in the field. Dave knew all the tricks or things to try. In
many cases, the things we tried didn t work, but each gene that you work with is
different, and that s why you can never predict success, and that s why I think the
patents are important. When we were trying to defend these cases in court, the patent
attorneys would say, "Well, it s obvious that it can be cloned." And my response to
them was, it s not obvious that you would ever succeed. You can try, but it s not
obvious that it would work because every molecule is so different. And there s
problems with each project that we encountered. So you have to try different strategies.
With t-PA, I tried five or six different strategies just to get the 5-prime end. There s not
a cookbook that tells you how to do certain things. There s things you can try, but
believe me, everything s a struggle.
Hughes:
Pennica:
Hughes:
Pennica:
I can imagine.
So basically, we published the first wild-type p53 clone, which was quite exciting, given
the fact that I had people saying, "Why are you working on p53? What is p53?" And
then later, I would say, "Molecule of the Year," because we didn t know at the time.
Do you want to bring home the importance of the fact that you were working on the
wild-type? Is it just simply that most people were working on a mutant?
The fact that most people were working on a mutant, they thought it was just involved
in tumor growth. In fact, they were misled because they thought that was p53 s
function, where in fact, the wild-type gene acts to protect, acts as a tumor suppressor,
acts to protect the DNA or the cell from damage. So it s actually a three hundred and
sixty degree difference in function, based on the fact that others were working with the
mutant forms because they were isolating them from tumors or tumor cell lines which
had made mutations.
Hughes: Is this sort of thing a worry in the back of scientists minds, that maybe they are
working on the wrong version of the molecule or the wrong virus? I mean, I m thinking
also of the development of the AIDS vaccine, where there was concern at one stage that
the laboratory strain of HIV was not the one that was circulating in the population,
particularly in Africa. All this work had been done on this particular lab strain that
might not be relevant to the real world.
Pennica: Right. 1 think that s a worry that every scientist has, and you have to make sure you re
not working with an alternative splice form. We now know that one base change can
change the function of a molecule. So now we have methods where you can screen
populations to look for polymorphisms or changes and different splice forms. The
computer programs we have now can analyze a gene and pull out different splice forms,
38
so it s easier to look at the different splice variants or polymorphisms than it was twenty
years ago. Now that we have these programs available.
Hughes: And would that be one of the first things that you would do in starting a new project?
Pennica: You first have to clone the molecule. If it s already cloned and known, then yes. Then
you can look to see what the variants are. But you don t know the function of these
variants. So a lot of the projects that we ve worked on since then have shorter 3-prime
ends or 5-prime ends or missing chunks in the middle, and you say, "What do these do?
Why are they important? Are they important?" It takes years of work to figure out
whether one that might be thirty amino acids shorter at the 5-prime end is relevant, or
has anything to do with making a tumor or preventing a tumor from forming. It s critical
to know that they exist, but then it takes a lot of time to figure out whether those
differences are important.
Hughes: And what about actually obtaining the molecule? If it s not derived from a cell line that
is already in Genentech s possession? Is that something you get from literature, to go
back and see who s working on this?
Pennica: If it s been cloned, you mean?
Hughes: Yes. Or not cloned, too, if you re just beginning a project, and you need the protein.
Pennica: Well, we try to isolate it. I mean, that s the thing that
Hughes: So you would do that here.
Pennica: Yes. We would try to find a cell line. The first thing you d do is either try to find a cell
line that makes a lot of the protein or a tissue that expresses a lot of it. So you d grind up
a heart, or a liver, or a lung where you think it might be made and try to purify it. Again,
that wasn t my expertise. The protein chemists would do that. And they would get it
down to a single protein. We wanted something that had an activity, so we would assay
for it. With t-PA the assay was to see if the protein dissolved artificial clots in a test
tube. With TNF, the protein killed cells in a petri dish, so that s a nice assay. But if you
don t have an assay, if you don t have some function, then it s ten times harder. So you
know that there s a molecule out there. You have to have some way of identifying it
because how do you know you ve ever purified it, how do you know the function?
That s why I said every protein that we ve worked with, or every cloning project that
we ve done, has been a challenge because there s always something different that you
have to worry about.
Hughes: It s interesting, isn t it, that because of the way, at least the media treats it, and maybe
even and I should ask you this in the way the paper s written up: It s the cloning and
the expression that become the big things. Those are the triumphs. The sequence of
events has to be there in order to get to that particular stage, but somehow, and I think
it s understandable, because I suppose that s considered fairly routine and unexciting.
The exciting part is the cloning and the expression, am I right there? These other stages
kind of drop out of the story that s being told.
Pcnnica: Back then, it was the thing to clone and express a new molecule. You all of a sudden
had something that killed tumor cells, for example, for TNF. That was very exciting.
Nobody knew what the protein looked, like. That to me is so exciting. What does the
molecule look like? Does it have anything similar to other molecules? And there had
been so few molecules cloned that you couldn t compare it to anything. And now it s
not enough. You can t just publish a cloning paper unless it s something very bizarre
that someone s been working on for a hundred years, twenty-five years, thirty years,
that all of a sudden they clone. But now, the journals want mechanism. They want to
know what this molecule does. They want to know what role in the body it has, and so
you have to do a lot more than you had to back then. Although, back then, given the
technology and the time, it was a huge amount of work. So just because things have
progressed now, people clone things in a day, where it took years. It s a whole different
set of criteria for what the journals will accept these days.
Hughes: And that must have meant changes in Genentech s organization, too, or at least beefing
up certain parts of the organization. What I hear you saying, and correct me if I m
wrong, is it s not just enough to identify, express the molecule: You have to start on the
way to showing function.
Pennica: Exactly.
Hughes: So the people who are showing function aren t the same people, necessarily, who are
doing the cloning, right?
Pennica: Exactly.
Hughes: So then you would have to hire all those people who are into the biology and the
functional part of it.
Pennica: Initially, 1 was doing the cell-killing assays, and then when we were doing hundreds and
hundreds, we d give it to another group who would do it in 96-well format, and then
they would give us back the activity data. A whole other group would optimize, make
sure everything was perfect, and give us the information back so that we could continue
cloning. So all of those people were critical. So it was very easy to determine who went
on a publication, because you could not do it without these people.
Hughes: And does it work that way today?
Pennica: It still does work like that today, especially, because we have an animal group that does
all the tumor studies. We have an immunology group that makes the antibodies. We
have assay groups. You cannot work alone, isolated. We tried way back in 1 980; we
had to be jack-of-all-trades. So I was doing a little bit of protein chemistry, a little bit of
assay work. But today it s much more efficient and everything moves much faster by
giving it to the experts. "Do this for us. Will you please help us with this? Do this." It s
a well-oiled machine here now.
Hughes: But is the science, perhaps, for the individual less interesting, because you are doing
you re using the same approach time after time rather than being a jack-of-all-trades?
40
Pennica: Well, for me, I found it more stimulating in the early days, 1 think, because I was
learning; I was doing different things, and I had to learn how to use an HPLC machine
and an FPLC machine. Frustrating, but I was able to do it, but each day was something
new and different, and it was a whole different environment back then, where if you
needed to do something, you learned it. You had to do it yourself because there wasn t
enough people here to do it, and 1 enjoyed that. And now you hand it off. So it doesn t
feel like you have as big a piece of the pie, 1 guess. But you are in charge here. Again, I
have charge of five projects that I m working on, and so I m the principal investigator,
but it s different. It s a different feeling for me at least.
Hughes: I suppose it s what inevitably happens with the evolution and success of a company.
Pennica: Oh, sure. And that s a good thing. I guess it s good because we can do things faster and
quicker than most people and that s why we have been successful. But for the individual
scientist, it s a different world here now than it was twenty-three years ago.
Hughes: And you probably lose people as a result of it.
Pennica: Oh, sure.
Hughes: The people that don t work so well in the compartmentalized environment. Where they
go, I m not quite sure because even academia, academic science, is more that way, isn t
it?
Pennica: Right. I ve been so fortunate because I ve worked on so many incredibly exciting
projects. I can t believe how lucky I ve been to work on t-PA and TNF and urokinase
and a few others I haven t talked to you about yet. But that is what has kept me here for
so many years, because it is always stimulating. Now we have a group that determines
whether there are proteins that are up-regulated in tumors, and so then we pick what we
want to work on, but those proteins have already been discovered. What we do is try to
push the experiments along to see whether you can make an antibody against that
protein which will kill tumors in mice. That would be a wonderful thing. But again, it s
not as much discovery for me. I found it a lot more exciting in the early days because
you were discovering new proteins. Today the proteins we re working on are known.
The exciting part now is: Will what I m working on become a cancer drug?
Hughes: Yes, I see.
Pennica: A different focus, I guess.
Hughes: You re further along in the process.
Pennica: We re further along, right. Molecules may need to be cloned, but again, we say to a
group, "Clone this." Three days later they come back, "Here s your clone." You know,
it s a whole different thing. It s not the exciting thing of getting the sequence back and
saying, "Oh my God, that s what it looks like." It s a whole different thing. But I had a
lot of those successes, more than most people.
Hughes: Have you had opportunities to go elsewhere and looked at them with any seriousness?
41
Pcnnica: I ve never looked seriously. I ve gotten many, many calls. But I always say, "I m happy
here." I think it would be hard to start over somewhere else. It would be very difficult
because it has been exciting, and I caasee the progress that has been made here. It
would be tough for me to leave.
Hughes: Well, it s good that you re happy.
Pennica: Yes.
Hughes: Anything more on p53?
Pennica: 1 don t think so. I think, again, there s a lot of effort and interest, and they have an
annual meeting, p53 meeting, every year.
Hughes: Do you go?
Pennica: 1 went once to the meeting, just because I wanted to see the progress that had been
made. It was about five years ago. Quite exciting because Amie was there, and he
would introduce me, "She s the woman who cloned p53." That was exciting, too. Many
people are still working on it. Whether it s going to be a drug someday, again, that s
still speculation. But now they at least know that this mutated protein is found in half of
all tumors.
Hughes: I gather that Genentech is not working on it anymore?
Pennica: We re not, no.
Hughes: Why was that decision made?
Pennica: Well, we were helping out Arnie, and at the time we were working on it, there didn t
seem to be any clear drug at the end.
I didn t know the patent situation when we were working on this. I don t know if Arnie
ever wrote a patent on this. He may not have. That I don t know. I m certain, well not
certain, but I m pretty sure Genentech did not write a patent on this. We were just
helping him out at the time. Dave said, "You want to work on this?" I said, "Sure."
Hughes: Do you ever say no?
Pennica: No, I never said no because it was always exciting, something new. I learned something
from every project that I did, and it was so much fun it s hard for me to describe how
much fun it was.
Hughes: After the t-PA project, when, understandably, you needed more direction from Dave
presumably than you needed afterwards because you hadn t cloned, did he more or less
drop out of the picture in subsequent projects when you knew how to clone?
Pennica: When I knew how to clone, he would be there for advice. He had less of a role than he
had in the beginning of the t-PA project, where it was every day showing him data, and
we would decide what the next strategy was. So as I learned, he had less and less
42
influence. I d see him once a week or show him my data. He had a vision and an insight
that was terrific.
Hughes: I m interpreting, but from what you told me last time, I got the picture of you, at least in
those early days, as working pretty intensely and independently. I mean, I think you
said something about not being in the social swim, for a variety of reasons, but mainly
because your nose was in the science. In contrast, 1 get the feeling in those very early
days about the maybe up until the time that you came that the handful of scientists
that were here were really talking to each other all the time. Consequently I m
wondering, do you think you ever missed out? I m not thinking so much of the social
aspects but just the scientific exchange that might have occurred if, as I m picturing it,
the main person you were interacting with was Dave.
Pennica: I probably did miss out because I was totally bench-focused, so to speak. I had a goal,
and I was known as not socializing and going around and talking about what other
people were doing, except with the people who were directly involved in the projects
that I was working on: Gordon Vehar, Bill Kohr, and Dave. But I didn t go around and
find out what Axel Ullrich s lab was working on or Peter Seeburg. I just didn t. I didn t
have time. The lab work is so time-consuming. It doesn t lend itself to walking away
because I usually had four things going at once.
Hughes: Were you an exception that way? Do you think the others were walking around to other
people s labs more than you were?
Pennica: I think so. Dave probably not.
Hughes: Well, and Axel impresses me as a pretty science-driven person as well.
Pennica: Yes, he is, and he probably was, too, in the lab all the time. But again, he was on a
different floor so I didn t interact with him. I knew who he was. But I didn t find out
what he was working on. So I sort of had blinders on, and I continue to be like that. It s
good and bad for me. [laughs]
Hughes: You get to the goal.
Pennica: Right. True. And I think it helps me get to the goal quicker than the competition because
there are a lot of people who are no longer here who would walk around and talk to
everybody, and they weren t as interested in doing the lab work. But I loved it. I just
had so much fun because it usually worked. It worked for me, and it was a fun thing to
do, and 1 miss it. But it s too hard to go back.
Hughes: Well, what about the cardiotrophins? Should we talk about them next?
Pennica: Yes. I need to break, [tape interruption]
Hughes: Diane, I think you were going to say something about uromodulin.
Pennica: So in 1987, we embarked on a project because some researchers had discovered an
activity that they had isolated from the urine of pregnant women that was reported to be
a potent immunosuppressant molecule. They found that it inhibited antigen-induced T-
43
cell proliferation and monocyte cytotoxicity in vitro, at very low concentrations, and
they also had shown that it was a high-affinity ligand for interleukin-1, as well as a
number of other immunosuppressive activities. The reason I wanted to talk about this is
because there are some projects in science that you work on for a long, long time, and
they don t turn out to be that spectacular. But you do learn something, no matter what
you work on, and this is something that I always tell the people who work for me: That
even though you may work on something that may never go anywhere, you learn
something, and you learn how to approach the next project.
So this sounded like a very exciting project, uromodulin, and we went about cloning
this protein by conventional means, once again getting protein sequence and screening
the library, and found that it was a protein of 616 amino acids that turned out to be
identical to the Tamm-Horsfall urinary glycoprotein, which was the most abundant
protein in normal human urine. So, in fact, that is a little bit amusing, but we did leam
something. Tamm-Horsfall glycoprotein had not been cloned before, but the amino acid
composition was identical, and we were able to deduce the fact that what we had cloned
was, in fact, this protein. We were fooled because it wasn t a pregnancy-specific protein
at all. It took a lot of work and not all things pan out, but basically, sometimes, things
like that happen in science. And again, I don t know what has been shown in the
literature as to the function of uromodulin or Tamm-Horsfall urinary glycoprotein
because I haven t followed it. We dropped it after this. That was basically the end of the
project.
Hughes: And you dropped it because it was the most prevalent.
Pennica: Yes. In fact, it wasn t a challenging cloning project once we knew what it was. But
when we collected urine samples from males, females, and pregnant females, we found
that the protein was the same in all those samples.
Hughes: Not too diagnostic for pregnancy then.
Pennica: Not at all. [laughter]
Hughes: I think there s also humor in science.
Pennica: Yes, that s true.
Hughes: Oh, good. All right. Well, shall we stop for the day?
Pennica: I think so.
Hughes: And carry on the story next time?
Pennica: Yes.
Hughes: Yes.
Pennica: Because there s a couple stories: the CT-1 story
Hughes: Shall I turn this off?
44
Pennica: Yes.
[End of session]
45
INTERVIEW 3: AUGUST 6, 2003
[Tape 4, Side A]
liughes: We ll start today with the history of cardiotropin 1?
Pennica: Yes, and what I thought I d do is give you a little bit of background to explain why we
got excited about this project. It s still exciting because I have collaborators now who
I m dealing with all over the world three manuscripts, in fact, on my desk right now
from scientists who are still working on it. So maybe one of these people will discover
something exciting.
We discovered a new molecule, which is so exciting to me. Just to begin, to tell you a
little background, we were interested in cardiovascular disease because it s the number-
one killer in the United States and Europe. Unfortunately, every thirty-four seconds a
person dies of cardiovascular disease, and there s over a million deaths in the United
States alone. Over six million patients in the US and Europe have been diagnosed with
congestive heart failure, which I think is terrible. Interestingly, the one clinical finding
in people who have heart disease or who are dying of chronic heart failure is that they
have an enlarged heart, which was puzzling, initially. This excessive enlargement of the
heart is known as cardiac hypertrophy, and this results in an increase in the size of the
heart muscle cells themselves, but not an increase in the number of cells. So basically
the heart is stretching.
To tell you a little bit about some of the features of congestive heart failure, you can
have damage from a heart attack or valve disease. The ventricle dilates and changes
shape, and this is caused by hypertrophy of the cardiac myocytes. The myocytes are the
individual cells that make up the heart, and the heart doesn t pump efficiently. So what
happens is that you get excess fluid that builds up in your lungs, and that s called
congestive heart failure.
At the time we started working on this, the mechanisms involved in hypertrophy were
relatively unknown. They didn t know what molecules cause this, why this is
happening, but it usually resulted from damage to the heart muscles themselves. There s
something called left-ventricular hypertrophy, and it s a powerful risk factor for
coronary heart disease and congestive heart failure, and it s usually associated with
being male and with advancing age, interestingly enough. The most common cause of
hypertrophy is high blood pressure, so you have to watch your blood pressure. This is a
fascinating picture, which shows cardiac hypertrophy caused by high blood pressure.
The normal heart is about 2 1 3 grams, and a hypertrophied heart is almost twice the size,
so it just stretches. This is an amazing picture that was published in 1949 to show that
they knew about this way back then, but they didn t know what caused it.
There s also something called exercise-induced hypertrophy, which I thought was
fascinating. You have a normal heart muscle cell, called a cardiac myocyte, and if you
do isotonic exercises, like running, swimming and basketball, what happens and this
is quite interesting is that the myocyte length increases, and there s an increase in the
diastolic volume in your left ventricle. If you do isometric exercises, like weight lifting
and wrestling, on the other hand, the diameter increases. And this is something called
pressure overload. The pulmonary artery, or your aorta, becomes constricted, and what
46
happens is that your heart wall increases in thickness, so you get two different types of
hypertrophy. You get stretching or you get an increase in the size.
This was all new to me when I started this project, so 1 thought it was fascinating. Then
I looked into trying to figure out: Why is hypertrophy induced? You have adult cardiac
myocytes, and these are terminally differentiated cells. They stop dividing shortly after
you re born, and in response to any stress or certain stimuli, like exercising or high
blood pressure, you have some heart muscle injury. Any demand for increased work by
the heart, which can include exercise, the heart cells adapt to this increased pressure or
workload by activating what s called a hypertrophic process. It s initially
compensatory, but it can become pathological. It can go through a transition where it
becomes pathological. We were interested in what factors turn on this process, going
from compensatory to pathological, and are there different hypertrophy factors for the
different stimuli. For example, if you exercise, is one protein made to make that length
increase in the myocyte? If you have a heart attack, is there another factor that s made?
We wanted to know what protein caused this.
Hughes: And is "we" you and your RAs?
Pennica: I was working with William Wood at the time, and this was a project I started shortly
after Dave left, [pause]
So I didn t want to study the factors that are known. Some of the genes are induced and
increased during the hypertrophic factor process, but we were interested in finding
something that was new. We developed a hypertrophy assay that was quite unique,
which I though was quite classy at the time. We took newborn rat hearts and isolated the
heart cells from these hearts. We put them in 96-well dishes, these little plates that have
ninety-six wells. You may have seen them.
Hughes: Yes.
Pennica: We waited twenty-four hours. You add the substance you think causes hypertrophy, and
you stain the cells with a dye so you can see them clearly under a microscope, and you
score them visually to see if they have stretched. It was a really simple assay. You can
test substances to see if the cells go from looking all rounded, to stretched.
Hughes: I see.
Pennica: Which is quite amazing. And so we had an assay. Just to show you what it looks like,
we scored them based on the size of the cardiac myocytes in a dish. Normal cells are all
small; they look like regular cells, but we knew some compounds that would induce
hypertrophy, like phenylephrine. We scored them on a score from three to seven, where
seven was maximum hypertrophy, the cells were as big as they re going to get, and
three was normal-sized cells.
Hughes: This is just looking under the light microscope?
Pennica: Yes. They re purple because we stained them with crystal violet. So we thought, great,
we have a wonderful assay where we potentially discover unknown hypertrophy
factors. Anything that might be known, we can put on these cells, and see the response.
47
This was a heroic effort by a woman named Kathy King, who spent hours, days,
months, looking under a microscope, to look for subtle, subtle changes because it s not
that dramatic. It would be great if it would go from small to big, but in some cases it
went from here to here, and she was able to see a difference. If it weren t for this
woman, working so diligently on this assay that s why you need the people to do
work like this 1 couldn t have cloned CT-1. It was just tremendous that she did this.
So we tested a number of factors, and like 1 said, phenylephrine was one, and there was
another compound called L1F, leukemia inhibitory factor, which will become important
later on, that seemed to cause this change from no stretching to fully stretched and
extended cells. So we had an assay and that s one thing for any cloning project: You
need to know what you re testing for so we had an assay, the cells get big, that was
easy to see. I mean, that mimicked what occurred in your heart. People knew that
there s a cell-type called embryoid bodies that we decided to use as a source to try to
find novel hypertrophy factors. Initially, these are mouse embryonic stem cells. You ve
heard a lot about embryonic stem cells, and we took embryonic stem cells that are
normally undifferentiated. What happens if you normally plate them on a layer of other
cells called fibroblasts and you can also grow them in the presence of LIF, or
leukemia inhibitory factor? What happens is: If you remove the fibroblast feeder layer,
or you remove LIF, the ES cells differentiate. They divide, and they become little
embryoid bodies, little round circular balls, that, interestingly, spontaneously beat. So in
a dish, you can see these little things pulsing. It s amazing. They also display heart-
specific markers, or proteins, that are produced, and they re a source of ventricular
myocytes. You can actually isolate ventricular myocytes from these little embryoid
bodies. Again, they re microscopic. You could see in a petri dish all these little specks,
and you look under the microscope, this is exactly what they look like, little balls. So,
we thought, great, we have an assay. We have a source of factors, unknown things that
people had not yet discovered the fact that they beat, they re mimicking what s
happening in the heart. So maybe there is something that the embryoid bodies produce
that can make the heart cells stretch. That was our hypothesis; this is what we were
guessing. So we got lucky. This was such an exciting project for me.
We did find that hypertrophy was induced by the embryoid body-conditioned media.
The conditioned media is whatever the cells are sitting in at the time. It s just like a
broth or a soup. We would take media that did not contain embryoid bodies, put it on
cells and stain them, and the cells looked the same, and then we took media that these
little embryoid bodies had been cooking in for twenty-four to forty-eight hours, and the
cells got big. So this was exciting because we thought, great, we have a source. We have
a source of factors that cause hypertrophy. We didn t know what it was because there
are some known substances that will do this that had been published in the literature.
Here s a beautiful picture just to show you what the stretched cells look like. You can
stain for a marker called atrial naturetic peptide, which is this yellow color surrounding
the nucleus, and you can see, these are called sarcomeric units. The cells stretch, look
like little train tracks. But this is what happens to your heart when they re stretched like
that or when you exercise. So we had an assay, we had a source, and now we decided to
do an expression cloning project where we would take these embryoid bodies and the
media am I making this clear so far?
Hughes: Yes, you are.
48
Pennica: Great. I thought this was so much fun. We took these embryoid bodies and incubated
them until the media gave a hypertrophy score of seven, which is the maximum
stretching that you can get. Therefore, we knew that they were secreting a hypertrophy
factor. We extracted RNA from these embryoid bodies and made a cDNA expression
library, transfected pools of clones into 293 cells, which are human embryonic kidney
cells, so we had pools that we put on separate dishes. Each pool contained different
clones, and we incubated them for four days without serum, and then we took the media
from these cells, put them onto cardiac myocytes, and we had Kathy King look at the
cells and see if she saw stretching of the cells. And again, we had pools of 40,000
clones. So any activity, unless it was very robust, was going to give you a minute,
minute signal.
Here was our strategy. We had ninety pools that contained anywhere from 1 0,000 to
15,000 individual clones, meaning individual messenger RNA, that could cause
hypertrophy. So one out of 15,000 may be the clone you were interested in. We didn t
get any positives, and the assay score was three. So she looked at hundreds of dishes
that we transfected. Then we had tested 300 pools where we used smaller-sized pools,
from 1,500 to 5,800 individual clones per pool. Again, no positives. At this point she s
looked at close to 400 wells under a microscope. Actually, more than that because we
repeated this many, many times. Again, the assay score was three, meaning no
hypertrophy. Then we made the pools even smaller, only 400 to 1,000 clones per pool,
and all of a sudden, in one of the pools that had 700 individual clones, she said, "I think
I see something, and I ll give it a score of four." Now, that could be just junk; it could
have been her eyes or the day or whatever, but we got one pool.
And then in another pool, where we even broke it down further, where we had seventy-
five to close to 200 clones per pool, she got another positive where she scored it a four.
Again, such a subtle change you almost cannot tell, but her eyes were so used to looking
at this. She said, "There is something there." So we had two pools, one that contained
700 clones and one that contained 190. We broke it down even further because you
want to get that one clone that has the activity. That s the goal, right? So this was so
exciting. When we had a pool size of 190 we took the smaller size pool; we thought
that would be easier to start with we got a hypertrophy score of four. We broke it
down to eighty clones per pool, all of a sudden it went to five; twenty, a little bit higher;
and when we had one clone she gave it a score of six. We got so excited because we
thought we have something here. This was the first expression cloning project I ve
done. And this was so exciting because we found something. Now, you get excited
initially, and you say to yourself, what if it s something known?
Hughes: Yes, yes.
Pennica: That s a drag because even though it sounds like a very straightforward project, this
took a long time, over a year, to get this far. We had to develop the assay; we had to
develop our embryoid bodies system. There were a lot of hit and misses here. It sounds
very smooth as you go through it, but we thought, great, we have something. So you
have to see if it s real. And so we took this purified clone. You can see here that in the
absence of media from this one clone, the cell is very rounded, but then when you put
the media from this one clone on the cells it is very stretched and beautiful. So we got
very excited.
49
So we called this molecule cardiotrophin or cardiac myocyte hypertrophic factor. When
we went to sequence the DNA from this clone to see if it was unique, in fact, it was
unique, which was so exciting because it meant we found something that nobody had
ever seen before in the world. Interestingly enough, it was related to it belonged to a
family which included L1F, which remember 1 mentioned earlier on, leukemia
inhibitory factor. The family also includes ciliary neurotrophic factor, and a few other
members, oncostatin M, that are very similar. But there was weak hornology, so we
knew we had a new member of this family, which was so exciting. So again, between
the other members of the family, 1L-6, interleukin-6, interleukin-1 1, CNTF, oncostatin
M, LIF, there was a very low hornology, about 20 percent hornology between CT-1 and
the other members of the family. It wasn t tremendous, but we knew it was good
enough to make CT- 1 a member of this family.
Hughes: That s structural hornology, right?
Pennica: It s amino acid sequence homology.
Hughes: But can you leap to the conclusion from that that there s going to be some functional
homology as well?
Pennica: Good question. We didn t know. Because we knew that nobody had ever tested these
other compounds to see if they caused hypertrophy before. So that s a great question,
and we had to do that. So here, we call these the LIF family of cytokines. I mentioned
all of these to you. And LIF was cloned in 1984.
Hughes: Not by Genentech.
Pennica: No. Not by Genentech. CNTF also in 84, oncostatin M in 86, IL-6 in 87, IL-1 1 in
1990, and then, ten years later from when the first family member was cloned, we
cloned CT-1 in 1994. And they re all roughly the same size, anywhere from about
twenty to twenty-five kilodaltons in size. These proteins are known as cytokines,
secreted proteins that seemed to act on other cells. That s the definition of a cytokine. A
lot had been studied about the other family members: LIF, CNTF, et cetera. They knew
that CTNF was involved in neuronal cell survival. They knew that these other family
members were involved in bone metabolism, blood vessels, hematopoietic cells. They
seemed to have multiple functions, and macrophage differentiation, ES cell
differentiation. So knowing the activity of the other family members, we thought to ask
what CT-1 does, to see if CT-1 has similar activities.
This is a picture of a structure of the family, showing different cytokines here in the
circles, and then the receptors that they bind to on the cell s surface in order to exert
their activity.
Hughes: This is the cell membrane?
Pennica: This is the cell membrane, yes. And interestingly, each one of these family members
uses a common receptor subunit called gp!30, and that is critical for its activity. So, to
answer your question, do other members of the LIF family induce cardiac hypertrophy?
Nobody had ever tested this before, so that was the first logical question, great question.
And when we did this, we found, in fact, that the other family members do induce
50
hypertrophy. CT-1 seemed to be the most robust although LIF was very potent as well.
Oncostatin M was potent, the other family members not as much. So that was
something new that we included in the paper that we published. As 1 mentioned,
because it has homology, we tested whether CT-1 functioned in other assays where LIF
has activity. But I don t want to go into that because all this data is included in the
paper. But just to tell you the background, we did find similar activities. Because CT-1
had so many different activities, we didn t think it could be used as a drug that you
could just inject or make an antibody that could be injected, to decrease the size of the
enlarged heart cells, because it had so many other activities in your body. You want to
make sure a drug just does one thing, otherwise you could hurt the patient.
But one collaborator I now have collaborators all over the world who are still working
on CT-1 he is in Marseille, France, and he found that CT-1 supported the long-term
survival of spinal cord motor neurons. If you can find a factor that can support the
survival of a motor neuron, it could be a therapeutic agent for things like ALS or spinal
muscular atrophies. So this was very exciting, the fact that we found CT-1 caused these
neurons to survive longer in culture than other factors that have been looked at before.
We published a paper in the journal Neuron, which was quite exciting, and we got the
cover photograph showing a motor neuron here in red, that CT- 1 has been added to
it s quite dramatic. I thought this was a beautiful picture. [Neuron, Vol. 17, Issue 1]
Hughes: Yes, isn t it though?
Pennica: It looks very different from a heart cell.
Hughes: Absolutely.
Pennica: CT-1 causes this activity. You can see the neuron starts out like this and then extends
branches out, and that s what CT-1 does. So we thought that this could be a
physiological motor neuron survival factor. Other investigators I m collaborating with
now are investigating neuronal properties. We made a knockout mouse where you
delete the CT- 1 gene and try to figure out what happens to the mouse. We have another
collaborator in Germany who has made a triple knockout, where you knock out three
genes at once. He s knocked out CT-1, LIF and CNTF, three of the family members,
and he s analyzing the mice right now, and I got a draft of a manuscript just a week ago
from them. So this is quite exciting for us.
At this point, we don t know the true function of CT-1. It s still being investigated.
From 1994, we re still working on this, and we know things that it does, but in the body
it may work in collaboration with other cytokines; it may work in collaboration with
other family members. We still don t know.
Hughes: When you collaborate with these different groups, is your function usually more or less
the same? Are you, for example, usually the cloners or is there a technology that is
usually yours, and then you rely on other people for aspects of the research?
Pennica: It depends. For a project like CT- 1 , where we already have the clone, we would have
people writing to us, saying, "I saw your exciting paper and I want to do x or y can
we have your clone? Can we have antibodies? Can we have protein?" And my thought
is, give them to everybody who requests them because I m not able to do everything. At
51
some point 1 move on to another project because Cienentech may have determined that
this is not worth pursuing at this time. It is interesting, but may not be a potential
therapeutic target. I use my expertise in that 1 tell the collaborators if there s other
assays that they need done, we will do it for them because we have a hypertrophy assay
here. We will give them protein we ve made protein for them. We can make more
antibody. We just let them do their thing, and 1 get papers in the mail saying, "We re
putting you as an author on these papers."
Hughes: Does the legal department enter into this process?
Pennica: Oh, sure. Yes. 1 can t send out anything unless the legal department approves, and we
have the patent on CT-1, which is great. So any discovery, I guess I m not exactly
sure how it s structured, but we send them a Material Transfer Agreement form where
we tell them that we would like them to tell us their results, and if there s anything
that s patentable to come out, we have to review their papers before they can publish
them.
Hughes: And is one of your arguments this is put very simplistically to the legal department:
This is an avenue of research that we probably will not be pursuing and, so, let them do
it and give us some of the credit.
Pennica: Yes, let them do it because one of these researchers is going to find something really
exciting. It just takes years. It may be something we never thought of.
Hughes: Is part of the Material Transfer Agreement that your name or a Genentech scientist s
names will be on the paper?
Pennica: Not necessarily. It s up to the researchers. They decide whether your contribution was
enough to warrant authorship, and in some cases I am not put on the papers. In other
cases, they feel they couldn t have done the work without the things I supplied them
with: proteins, antibodies or the clones, so I think in that case it s warranted. I help them
out as much as I can. I review the papers. We have a very good review system where
any papers that come from the outside or even inside papers go through an internal
review where we have two people scrutinize the paper and give a very critical review
before it gets sent out to the journal. This is very helpful because they can pick out
things that you may not have thought of, and hopefully will strengthen the paper, so it s
always a good thing.
Hughes: Is that review both scientific and legal?
Pennica: Yes. It is both scientific and legal because legal looks at it thinking is there an
invention here? Is there something new that we haven t thought of? And if there is, then
we write a patent.
Hughes: Right. Yes.
Pennica: And in most cases our patent usually covers what they have, what they are doing. In
many cases it s basic research because these are universities I m collaborating with, and
they re all over the world. That s fun and exciting. Many of these collaborators I ve
never met.
52
Hughes: How do you feel, Diane, when a project that you have been working on and really
scientifically engaged in, turns out not to have at least immediate therapeutic promise,
and therefore you have to drop it and move on?
Pennica: It s disappointing because you ve put so much time and effort into it. But every project
you do, you learn something, and I d never done an expression cloning project before. I
knew nothing about cardiac hypertrophy, so learning all about the process and what was
involved was exciting. So you move on. You move on to the next one and say, "Oh,
what can I leam next?" That s what is so exciting, and the fact that people are still
interested in this almost nine years after the protein was cloned is great. I think people
are still interested enough to study this protein. I dropped it, long ago, but they may find
something that may be a potential drug at some point. You can t get too disappointed.
You can t have every one be a t-PA. [laughter]
Hughes: Exactly. Was it your decision to drop the project?
Pennica: It was a consensus, because everyone thought this protein had too many different
activities. Since all the other family members have multiple activities, we tested CT-1 in
all of those assays as well, and found CT-1 does this, it does this, it does this, et cetera,
and we thought if we inject it into people it might have too many side effects.
[Tape 4, Side B]
Hughes: seminar of the morning is on WISP. And what does that stand for?
Pennica: Wnt-induced secreted protein.
Hughes: Wind?
Pennica: W-n-t. So I m going to tell you a little bit of the history and the background of how it
was started in my lab. We finished the CT-1 paper in 1994, and 1 did a number of things
between then. But I ll go to the most exciting one next. And this project began in 1997
as a collaboration with Arnie Levine, and if you remember, Arnie Levine was the
collaborator who I did the p53 project with, way back.
Hughes: Yes.
Pennica: Arnie was on sabbatical at Genentech at the time, and we were between directors, so he
spent the summer here. He would review projects that we were working on. I came into
his office one day. He said, "I have a great project for you." I had shown him some of
the things I was doing to look for targets in stomach cancer, and he said, "I have a great,
exciting project for you." And that s all I needed to hear. Amie was always so bubbly
and excited about research that he could make anybody excited about a new project. He
said that we should look for colon tumor antigens that may be therapeutic targets using
the Wnt-1 signaling pathway as a cancer model.
Just to tell you a little bit about colon cancer to begin with, there are two key colon
cancer mutations. Eighty-five percent of colon tumors contain mutations in a gene
called the APC tumor-suppressor gene, which stands for adenomatous polyposis coli.
That s why people call it APC. Forty-eight percent of colon tumors that lack APC
53
mutations contain mutations in an oncogene called beta-catcnin. So our rationale, just to
give you a little bit of background about the pathway for using the Wnt-signaling
pathway as a cancer model, is that: Many, many years ago, Harold Varmus s lab
determined that Wnt-1 was an oncogene, meaning a tumor-causing gene, and if you
ectopically express it in mice, they develop mammary tumors. So the Wnt pathway
mimics what occurs in the majority of human colon tumors as a result of an APC or
beta-catenin mutation.
In a tumor cell, that either has a mutation in APC, has a mutation in beta-catenin, or if
you have Wnt-signaling, beta-catenin levels go up, the half-life goes to four hours.
Normally, beta-catenin levels sit around in the cell, and it gets degraded very rapidly
and has a half-life of about thirty minutes. But when you have a mutation in a tumor
cell, the beta-catenin molecule does not get degraded. What happens is that beta-catenin
protein increases in the cell, it gets shoved into the nucleus by some mechanism, and it
binds to transcription factors and turns on genes that are thought to play a role in growth
control and tumor progression. So these two mutations cause the same thing. You can
either have a mutation in APC, a mutation in beta-catenin, or Wnt-signaling. The end
result is the same: Beta-catenin levels go up, it gets shoved into the nucleus, genes are
turned on, and you get a tumor.
Why? What genes are turned on? So just to tell you a little bit about what Wnt does in a
cell, we needed an assay, just like we did for the CT-1 project. We needed a cell line
that potentially made a protein that would induce this tumor could be hundreds of
proteins. But we were looking for something that would cause this change. So we took a
mouse mammary epithelial cell line called a C57MG cell, and we put the Wnt gene in
these cells. When you did that, the cells changed shape, and they became elongated,
similar to what happened in the CT-1 project, and they lost contact inhibition. Normal
cells in a dish, once they touch another, would stop growing. So the loss of contact
inhibition meant that the cells would pile up on one another, and they became
elongated. By eye, it was a quite dramatic change from a normal cell to an elongated
cell once you put Wnt in the cells. When the cells are not transformed, we can also look
at the beta-catenin levels. In the normal cell, beta-catenin levels are very low, 1
mentioned, to begin with. Once you have Wnt in these cells, you get a tremendous
amount of beta-catenin being over-expressed.
So we had a cell line, then, that we could use to find proteins, again just like the CT-1
project, because these cells mimicked a tumor cell. And we thought, we can look for
proteins that were unique, [tape interruption]
Suppressive subtractive hybridization, basically the key word there is subtraction. We
had the parent cell line that did not express Wnt, and we had a cell line that we had
over-expressed Wnt in, and we did a subtraction. So we looked for all the genes we
did it basically in an array. Not to get too technical, we did an array of what genes are
different. You subtract out everything that s the same. You look at all the genes that are
different, and we got hundreds of genes that are different. When you do this cloning,
you end up getting a library of clones that are just partial pieces of genes. We ended up
sequencing inserts from 2,000 clones, many of which were known genes, many of
which were unknown genes. What I ended up doing over a period of many, many
months, because it took months to sequence these 2,000 clones, was to look for proteins
that might be unique and that might be related to other genes. I saw in one of these little
54
snips of DNA it was only seventy base pairs long but when we got the amino acid
sequence for that stretch, there were, 1 think, three or four cysteines that appeared to line
up with the cysteines in a family called the CCN family of growth factors. And so we
had, once again, isolated a brand-new member based on seeing the sequence from that
one small fragment of DNA. We had hundreds of others. There s other things that we
can still look at, but it turned out to be a brand-new member of this family, which was
quite exciting. Because it was novel, we could name it something new. So again,
another exciting project.
The CCN family of growth factors contained three members, connective tissue growth
factor, CYR61 and Nov, and we isolated three unknown proteins, which we named
WISP-1, W1SP-2 and WISP-3 because they were related. They were cysteine-rich
secreted proteins; cysteine-rich meaning they contained thirty-eight cysteine residues,
which made it very difficult to purify these proteins. It still is difficult. The exciting
thing about this family, since we knew nothing about WISP-1, is that when we looked
in the literature, very, very little was known. There were only about thirty papers
published. Now there are hundreds of papers that have been written on this family since
we started this project. We were looking for something involved in tumor formation. I
knew nothing about any of these family members. Again, that s why it was exciting.
We found out that these members were involved in cell proliferation. That s important
in cancer. They were involved in attachment and migration, which is important in
metastasis. They were involved in wound healing, differentiation, and angiogenesis,
meaning the infiltration of a tumor by blood vessels, and some of them in fact were
implicated in tumorogenesis. And so we got very excited, because we thought WISP-1,
WISP-2 and WISP-3 could also play a role in tumor genesis. So, we may have found
three new proteins involved in tumor formation.
So we did a literature search, and we found out what all three of these family members
did that could be involved in tumor formation. We found that CYR61 was up-regulated
in pancreatic cancer and breast tumors. Connective tissue growth factor was found to be
elevated in melanomas and sarcomas. Other people had looked at the protein called Nov
and found it in a very rare childhood tumor called Wilms tumors. They found elevated
levels of this protein. A lot of these family members have been implicated in cancer. We
thought, great, we have three brand-new members that might be implicated in cancer.
We just had to find out their function. We decided to focus on WISP-1 first.
We characterized WISP- 1 , we sequenced it, we looked at its structure and found it had a
fairly high homology to the other family members, between 35 and 45 percent
homology. So it was higher homology than the CT- 1 project I talked about earlier.
WISP-1 had thirty-eight cysteines. It s a bigger protein; it s forty kilodaltons. Since our
focus was colon cancer, we decided to look in human colon cancer cell lines to look to
see whether the protein is elevated. When we did that, we found that in fact in a number
of cell lines, WISP-1 was elevated. We also looked at the DNA copy number and that
we do to determine whether the gene is amplified. When a gene is amplified, this can
cause problems in the cell as well. So we took a number of primary human colon tumors
and found that, in fact, in the majority of them, the DNA copy number was about two
fold in 60 percent of the primary human colon tumors that we looked at. It wasn t very
dramatic, but it was at least two-fold. People sometimes don t get excited about two
fold, but I remind them that if your temperature is elevated two-fold, you die.
55
Hughes: Yes. Good analogy.
Pennica: Another thing that you have to do in any project is determine once you find out that
this might be a target for cancer, you have to find out where else in your body it s made.
And if it s made everywhere in normal tissue, that s not a good thing because it means
that it probably has a role and a function there, so if you try to inhibit its activity you
could mess up something critical.
It was expressed in a number of normal human tissues, but it was significantly over-
expressed, or elevated, in human colon tumors. About 84 percent of human colon
tumors we looked at had elevated levels of WISP- 1. So you can imagine a localized
target. We did not find it in the normal colon, but we found it very highly expressed in
colon tumors. As you can see, these little spots are the expression in a colon tumor
versus a normal colon, where there s no expression here. So that was quite exciting. We
did a number of other assays to determine what it does.
We are now in the process I won t go through any of the other details of the science
because that can be found in the publication but we re now in the process of writing a
paper to see whether WISP-1 might be involved in metastasis. It may play a role there
so we re quite excited. Another lab at Genentech has taken over that project, and they
are working on that right now as we speak. We don t know what kind of target WISP-1
might be, but it s still quite exciting because it s still high-profile at Genentech.
Hughes: And your lab still has a role?
Pennica: I act as an advisor, and we do assays for them. If they need things done, we help them
out, or we give them reagents and cell lines. I got this group excited about this protein
and they are taking it over because I have five things I m working on now.
Hughes: And that s all right with you?
Pennica: It is.
Hughes: You don t feel proprietary?
Pennica: No, because they ll include me on the publications so it s great. It s better that I learn
something new and move on.
So anyway, this was quite an exciting paper, and it still could play a critical role in
tumor formation. But that was our goal, just to find a downstream target in the Wnt
pathway that might play a role in tumor formation. We might have found something.
The fact that we found something unique is quite exciting. I ve been very lucky to
discover so many unknown proteins that had never been cloned before. That was fun for
me.
Hughes: Well, does that satisfy you in terms of what we have said so far, what you have said so
far, to cover at least the highlights of the science that you ve been involved with?
Pennica: Yes.
56
Hughes: Maybe you could make a comment about the technology and how that has impacted
both the efficiency and also the direction of the research that you ve done at Genentech
over, was it twenty-three years?
Pennica: Twenty-three years. Yes, the technology has changed dramatically. Back in 1980, we
had to do everything ourselves. For every project we worked on, if you needed
something done, you did it alone. We did work in teams; you couldn t do anything in
isolation. As I mentioned, Bill Kohr did all the protein sequencing because he had the
expertise to do that, but the cloning was so labor-intensive at the time. Now it is
cookbook. There are kits that you can use to do everything. The kits have become more
sophisticated, so in the WISP project 1 did use a kit, but it was still a little bit labor-
intensive but unbelievably easier than what we had done twenty years before.
Hughes: Give me an example of something specific that had changed.
Pennica: So you could make a cDNA library in a week in 1997, whereas back in 1980, it may
take months to make a cDNA library that you could analyze. Just the steps to make the
cDNA was time-consuming because we had to make all of the components. When you
have a kit, you just take out a little tube and you add it to the reaction. It was very
different twenty years ago. We had to make our own gels. That s another good example.
I was making my own gels, which often took an entire day, to make all the reagents, to
pour the gel, to have it set, to let it sit overnight, to use it. Now there are whole
companies dedicated to making gels. It s in a Ziploc bag, you tear it open, pop it in the
machine and load your sample. It takes thirty seconds. Unbelievable.
So things have changed dramatically, and now we have whole groups that clone what
we want. They have perfected the technique, and they can, in a few days, get you a
clone that you need. Everything is more streamlined. It s certainly a lot quicker, but I
think the questions are a little different that we are asking now. We re now trying to
determine the biology of these molecules to see if they re involved in tumor formation.
Back then, we were just identifying, not just, but we were identifying new molecules
that could be involved in heart disease and cancer. You have to learn all about the
biology of the molecule, also, to see if it s relevant or does anything of interest. Clearly,
it does.
Hughes: Obviously, you re describing a tremendous revolution in efficiency and just general
ease, but at the same time you are in a sense losing control in the fact that you have to
depend on a kit, or you have to depend on a sequencing group or
Pennica: Yes, absolutely.
Hughes: Did this require sort of a shift in your thinking about how you did research?
Pennica: Yes. When you rely on other people, you rely on their quality, their efficiency and
their we are fortunate at Genentech to have super-team people, who do incredible
work here. But if there s one kink or one person who just doesn t do their job
efficiently, then it can waste a lot of time. For example, Kathy King, who was a
superstar on the CT-1 project, without her expertise and painstaking patience to look
under the microscope at that hypertrophy assay, the project never would have gotten
done. So there are superstars, and then I have worked with other people who are not as
57
efficient, and it s very frustrating. When you do it yourself, you spend the time and
effort to do it right, but you have to rely on a lot of other people sometimes. They may
not work as quickly or efficiently as you do, but we re pretty lucky here. We have a
good team. It has changed, though; it certainly has changed.
Hughes: Do you have any feelings in general about the fragmentation of research?
Pennica: I think it makes us do things a lot quicker. I know it does. I liked having a project all to
myself like I did in the old days. Now I feel there are a lot of cooks, which is a good
thing and a bad thing. It s hard to explain. You are at the center of a project in the old
days, and now you re more of a team player.
Hughes: Yes, yes, I see. Which 1 think for you, because 1 pick up and tell me if you think I m
there that at least in those early days, you were a perfectionist. You had your way of
doing things and you weren t going to be interrupted by Tom Kiley or anybody else,
[laughter]
Pennica: Right. Right.
Hughes: So I m thinking that it probably did take quite a shift.
Pennica: It did.
Hughes: For you, as a person, to all of a sudden being reliant on somebody whose efficiency
well, whose perfectionism might not have been at quite the level that yours was.
Pennica: Right. It has helped me, and it also hurts me in a way because you get frustrated when
people don t have the same level of fanaticism as you do.
Hughes: Yes. Yes.
Pennica: I always think I am fanatic, but 1 think that s what has helped me be successful, the
fanaticism and attention to detail. Many people don t put that much time into doing
something and many times their experiments don t work. Well, my experiments worked
90 percent of the time because I was fanatic about them. It took me a lot longer to do
things than other people, but a lot of people left science because they didn t want to be
that fanatic. It takes a lot of patience, and you have to go through a lot of failures. But I
would do something thirty times until it worked. It may not work because you re adding
the wrong buffer, or you have too little or too much buffer, and you don t know what
it s supposed to be. Back in the old days. I would do an experiment forty times until it
worked. That s the way that I was taught when 1 was doing my undergraduate work, and
in graduate school, to try a hundred things until you do get it to work. So I didn t like to
give up.
Hughes: Do you try to pass on that approach to your laboratory people?
Pennica: I do, yes. I try to tell them when you re running a gel and you only have two samples,
throw in a few more samples because you never know, you might find something
exciting. Of course, it s easier to just run two samples, but if you have a chance to look
at five other DNA, RNA or protein mixtures that may be related, put them on the same
58
gel. It takes more time, it takes more effort, but that was my approach: Instead of doing
less, do more.
Hughes: And is that a characteristic, or an openness to that approach, that you look for when
you re taking people on into your lab?
Pennica: Well, it s hard to measure. You never know how much enthusiasm someone is going to
have. I try to find somebody with a spark always and who is ready to jump out of their
chair to do the research, so they usually want to do more than less.
Hughes: Good. Well, I have a few more general questions if you feel happy with what we said so
far.
Pennica: Sure.
Hughes: Do you have until twelve?
Pennica: Yes.
Hughes: As I see it, you ve lived through three regimes at Genentech, the Swanson, the [G Kirk]
Raab and the [Arthur] Levinson. I know from what you ve told me before that your
nose has been in the science, and I have the strong impression that you weren t as
interested in the larger picture at Genentech.
Pennica: Precisely.
Hughes: Nonetheless, it must have had some impact, and I m wondering if you could say
something about any shifts in general functioning of the company as a whole that you
might have noticed as a scientist as you moved from one CEO to another.
Pennica: Well, I ve been very fortunate in that I ve been given great projects. I ve worked on
tremendous projects, and Bob Swanson was always so excited about everything we
were working on. He wanted this company to succeed, and he made it happen. I
interacted a lot with Bob Swanson because we were such a small company at the time;
there were only sixty people. I interacted rarely with Kirk Raab because we were getting
much bigger at the time. I don t know how many people we had then, but he certainly
knew who I was. But he never came around to talk to me, so I didn t know him that
well. And Art Levinson, I have interacted with since I got here. He got here four days
after I did, and so I interacted with him as a scientist, so he knew me well. There s never
been a time when I wasn t excited about the science.
The management changes have been excellent, as far as I m concerned. Because I
didn t interact with Raab, I can t comment on how the company went at the time. 1 think
Art s doing an excellent job, and Bob Swanson also did a tremendous job. So clearly
we ve become successful, as evidenced by our stock going up, and all the drugs we re
making that are now helping to save lives. So I feel fortunate that I was a part of that in
the beginning, and it s continuing now. Hopefully, I can make more contributions, but
one is more than most people ever ask for or more than most scientists ask for.
Hughes: Well, in 1990, as you know, Roche acquired a controlling interest in your company.
59
Pennica: Yes.
Hughes: Did that make any difference?
Pennica: Not to me.
Hughes: Not to you.
Pennica: Not to me at all. 1 have been fortunate that the higher-ups usually leave me alone and let
me do the science. I don t get involved in the politics. I don t usually know half of
what s going on as far as the politics go here. But that s a good thing because 1 stay
focused on my research and that s my strength. The Roche takeover, when they
acquired part of us, 1 never noticed a thing. Nobody ever came in my lab and said, "You
have to switch working on what you re working on."
And when Arnie came, 1 was working on gastric tumors, and he suggested that 1 work
on colon cancer, and so I started working on the Wnt project. So for the longest time, I
was working on what 1 was excited about. Dave Goeddel had always said that, "You
can t make people work on something unless they re excited about it. Because if you
tell them what to do, they will not do a good job." So I ve always had exciting projects,
and 1 just delve in and leam everything 1 can about it, and try to do the best 1 can. That s
a good strategy. If somebody says, "Oh, I have no interest in working on this or that,"
then they re never going to do a good job. So it s a good strategy. I ve never said no;
I ve always enjoyed everything I ve worked on and learned a ton, a tremendous
amount.
Hughes: Do you have any philosophy about when, if ever, a scientist, or I guess in this case, you,
should address the ethical dimensions of the science that you or Genentech is engaged
in?
Pennica: I m not sure what you mean.
Hughes: Well, I guess it was most apparent in the beginning, even by the time you came in 1980,
it was beginning to die down. But you know, there was the recombinant DNA
controversy. Should scientists tinker with the genetic endowment of human beings? It
was quite a debate in the nation as a whole and also eventually internationally with
Asilomar and that whole history of events that went on.
Pennica: But 1 don t think people understood
[Tape 5, Side A]
Pennica: cloning human beings, again, that s been in the news these days. What we re doing is
we re trying to identify genes in your body that are important for a particular function,
for example: What causes cardiac hypertrophy? What causes colon cancer? These are
simple questions, and there s probably not simple answers. We clone one gene, and we
hope it may be the magic bullet, but it might not be, at least not in isolation. You may
have to use a combination of things. But when we say cloning, we mean trying to find
the piece of DNA in your body that is responsible for a particular function. And so you
isolate that gene, put it into a cell, and make a protein, and then can you use that protein,
60
like t-PA for example, to dissolve a blood clot. In fact, in the case of t-PA, a simple
protein was enough to dissolve blood clots. We tried to do the same thing for CT-1 . In
this case, find a gene that causes your heart cells to get bigger, or undergo hypertrophy.
Well, you wouldn t want to inject CT-1, because it would make the heart bigger, which
is not a good thing. So, what we would want is an antibody against it, to shrink the
enlarged heart cells.
If people understood that you re not putting anything foreign into your body, you re
putting in proteins that your body already makes, sometimes in short supply. For people
who have blood clots, clearly they don t have enough t-PA circulating in the body to
dissolve them, for whatever reason; it could be genetic. So you re trying to supplement
the lack of enough t-PA. Human growth hormone, or HGH, is the perfect example, for
children who have a growth deficiency. We wanted to isolate that gene and make
growth hormone so we could treat children who lack sufficient HGH in their bodies. So
to me, if people think of it that way, it s certainly a great thing. Perhaps enough people
hadn t explained it in these terms. I don t know. I just think there were too many people
against it who didn t understand its potential. I think it s the most tremendous thing
that s ever happened for disease treatment.
Hughes: There are always lines over which at least some parts of society think that you should
not go. I ll give you an example with the growth hormone. I think most people would
agree that using growth hormone for children that are severely inhibited in terms of the
growth pattern has got to be a good thing. But apparently, there was an effort at one
stage to convince the parents of a population of short children, but not dwarf children,
that to show whatever image a parent or this society I m not quite sure where these
images come from of the height that a child should reach, and you turn to growth
hormone for what would be, in essence, a more aesthetic use than a therapeutic use. It s
a very gray line where you go from one to another. That s one example of that comes to
mind where things might go too far.
Pennica: Oh, there s certainly going to be some unethical doctors who prescribe it for the wrong
reasons. We hope there s not many of those people. But clearly, that s not our intent.
Hughes: Yes. And it s not something that you worry about.
Pennica: Well, it s disturbing when you hear stories like this, but you hope that they re few and
far between. We re not going to hear many stories like that.
Hughes: In terms of Genentech s legacy that sounds very elegant and overwhelming.
Genentech was, of course, the first to base itself on recombinant DNA, a pioneer in the
field, and I m wondering what the upsides and the downsides are of being the first and
the foremost in a new area.
Pennica: I think there s only an upside. It s just like publishing a paper: You always want to be
the first. That was my strategy and so you worked ten times harder than everybody else
to be first because I didn t want to be beat on the cloning projects that 1 was working on.
But you never knew who was working on them. So being first as a company, 1 think, is
only a good thing. You set a standard, for sure, because the image could get tarnished,
but hopefully it has not so far. I think people continue to say that Genentech is the star
61
of the whole biotech business, and I think it s just a good thing. 1 feel very fortunate to
be here for twenty-three years.
Hughes: You said something off tape about how people react when you tell them that you re a
scientist, and I m wondering if you ll say more about that, and also is that how you
define yourself?
Pennica: Well, I ve had many people just say, "No way, you can t be a scientist." I don t fit the
profile. 1 say, "What is a scientist supposed to look like?" But when they ask me what I
do, I do say, "Scientist. 1 do cancer research." That s easy for most people to
understand. So I m very proud of the fact that I do research.
Hughes: And do you define yourself to yourself as a scientist? Is that what you are?
Pennica: Yes. 1 think so. I think it s a good description: You re trying to discover new things, and
in this case we re working on a lot of genes now that are known, but we re trying to see
whether these genes are involved in tumor formation. So that is also a discovery
process. It takes a long time just to work on one gene, to find out whether it plays a role,
or an important role, in tumor formation or tumor maintenance. That s what I m
working on now. 1 have five proteins that we re trying to figure out what role, if any,
they play in prostate cancer and ovarian cancer. So 1 say, "Yes, I am a scientist."
Because if I d say molecular biologist or cloner, people don t understand that. 1 think
scientist is easier.
Hughes: Well, at last count, and I m sure it s changing, you had thirty-seven patents on which
you were inventor. I ve counted them up.
Pennica: You did. [laughter]
Hughes: Well, I m sure there may be more pending. And say something about how you value
those patents in reference to your publications.
Pennica: Well, the patents are something necessary for Genentech. I think it s very prestigious if
you are an inventor on a patent application because it shows that you were involved in
the thought process of defining something new and novel and finding a new use for
something, or finding a new use for something that was already known. It s an honor to
be on a patent application. But those don t get published as readily. They get published,
certainly, but for the scientific world you re more known for your publications than
your patents.
Hughes: And so if you had to choose between them?
Pennica: Oh, the publications.
Hughes: I thought you were going to say that.
Pennica: Yes, absolutely.
Hughes: There s the scientist, [laughter]
62
Pennica: Yes.
Hughes: Comment on your role in various litigations. Are there stories to tell?
Pennica: One in particular was quite amusing. I don t know if 1 told you this or not; maybe I did.
I can t remember. But there was a litigation it was a patent hearing in Germany, and
they were questioning the validity of our t-PA patent, saying we didn t make an
invention. They claimed it was obvious anybody could have done it. And so 1 went with
Steve Raines and a few other Genentech legal people. It was an arbitration where there
were three patent examiners deciding the verdict. I can t remember it was Kabi, I
can t remember. No, it was not Kabi; it was Behringer Ingelheim.
Hughes: Oh, yes.
Pennica: I think so. So there was a group of lawyers on one side. It was an L-shaped table, and
the patent examiners were in the middle. The Behringer lawyers were facing us, and
there were three or four Genentech people presenting the case. I was the only woman in
the room. The patent examiners were asking questions, and we would answer. 1 didn t
speak at that point. The lawyers would answer the questions from the patent examiners.
I would whisper things into our attorney s ear just to make certain points that we never
knew if this would work, for example. It s not obvious that you would ever get the
clone. It s not obvious that you would ever find something. They were asking questions
to us as well as the other side. We were never introduced to the other side to begin with.
I don t know how this happened, but 1 decided during one of the breaks to introduce
myself to the head lawyer for Behringer as we walked outside. I said, "I want to
introduce myself. I m Diane Pennica." The lawyer said to me, and I ll never forget his
words, "You re Diane Pennica? I thought you were much, much older!" He said, "I
thought you were one of the administrative assistants for the attorneys." So that was
quite amusing to me. It was clear that the patent examiners were favoring Genentech,
and in fact we won that arbitration. The German lawyers got so upset that they weren t
getting their points across that they decided to switch to speaking German instead of
English, so we all had to put on headphones to listen to a translator translate the
questions and answers. It was so bizarre. They decided not to speak English anymore;
they decided to speak German. So that was an interesting event in my past.
We also went to a trial in London, where it was quite fascinating because all of the
lawyers, including the judge, wore these wigs, these long white wigs.
Hughes: Yes, I ve seen them.
Pennica: And the judge was at a table that was forty feet above everybody else, and it was just
like you see in the old movies: these long robes and the wigs. I wasn t cross-examined
for very long. I think I was only on the stand for twenty or thirty minutes, but they really
kept Dave Goeddel up there for a long time, so he was on the hot seat.
Hughes: And again, was it the novelty of the invention that was at question?
Pennica: Yes. Yes, it was the novelty of the invention. We had Paul Berg as an expert witness on
our side. They had James Watson, the Nobel Prize winner who discovered DNA, as an
expert witness on their side. We had George Stark as an expert witness for us as well.
63
Hughes: What was Watson s argument?
Pcnnica: He was at Cold Spring Harbor at the time. I believe his claim was that because people at
Cold Spring Harbor were doing the same thing, it was obvious to do and that it would
eventually work. Well, it s never obvious that any cloning project is ever going to work.
So that was an interesting trial. It was fun to see the way the trial progressed.
Then there was another trial in Washington, and I was on the stand for a while. We won
that case. They asked me what the important points were. Our lawyers wanted to know
how to end, and I suggested telling the jury the story about me meeting the first heart
attack patient, because it tells them the human aspect of the whole trial and that we re
trying to save lives. At that time we weren t a big company just trying to make millions
of dollars. We were trying to save lives. And that seemed to be very effective for the
jury. It was scary being up there.
Hughes: Did legal counsel prepare you before you went up on the stand?
Pennica: They did. And I made suggestions on things that I thought would be useful to put in, or
include. They seemed to like me as a witness. The lawyers liked me because they said
they needed sincere people who the jury could identify with. I don t think the other side
was too hard on me in Washington. They were harder on Goeddel. Maybe because they
saw me shaking.
Hughes: I think one has to acknowledge that probably gender and age and all those things are
factors. It s not just a matter of the legalities and the science.
Pennica: Absolutely. 1 think that makes a huge difference. I was lucky, by meeting Desire Collen
because that first meeting where I wasn t kicked out because they thought 1 was one of
the guys daughters.
Hughes: Yes, I know it. Amazing.
Pennica: That was an advantage.
Hughes: Well, time is flowing, so I will ask you one more question and that is: If you had to pick
one contribution of which you are most proud, what would it be?
Pennica: Oh, of course, t-PA. Absolutely. I probably worked as hard on all of the other projects
that I ve worked on because I ve been so fanatic about every one of them but
certainly t-PA s given me the most satisfaction. Meeting heart attack patients and
getting letters and cards and phone calls from people has just been so rewarding. I
mentioned before that there will never be another t-PA. It would be nice if there were
others. I feel very fortunate and very lucky to have been involved in something like that
and had a great group of people to work with, who helped: Bill Kohr, Dave Goeddel,
Gordon Vehar. They were all instrumental in the project, and Desire Collen as well. But
I definitely worked hardest on that project, for sure, and it would be hard to work that
intensely again.
Hughes: Yes. Well, is there any more you want to add?
64
Pennica: I don t think so at this point, other than maybe looking at my old t-PA notes and seeing
if there s something I missed or forgot to tell you.
Hughes: That sounds good. Well, I thank you.
[End of interview]
Transcribers: John Lynch, Christine Sinnott
Final Typist: Lea Barker
65
APPENDICES
A. Curriculum Vitae 67
B. Letter of congratulations from U.S. Senate Subcommittee on Education, Arts
and Humanities, and photo of "Inventor of the Year" team 87
C. Science in action: photos from lab notebooks at the time of the successful
cloning of t-PA in 1981 (captions by Dr. Pennica) 91
D. t-PA in action: angiogram of restored blood flow; blood clot being dissolved 94
E. News reports and patient/ family testimony about the efficacy of cloned t-PA 95
67
DIANE PKNN1C A
APPENDIX A
F.DUCATION: 1973 B.S. State University of New York, College at Fredonia
1977 Ph.D. University of Rhode Island
RESEARCH AND PROFESSIONAL EXPERIENCE
1971 - 1973 Undergraduate Fellowship in the Department of Biology, State University of New
York, College at Fredonia
1973 - 1976 Teaching Assistantship in the Department of Microbiology, University of Rhode
Island
1976 - 1977 Instructorship in the Department of Microbiology, University of Rhode Island
1978 - 1980 Postdoctoral Fellow, Roche Institute of Molecular Biology, Nutley, New Jersey
1980 - 1988 Research Scientist, Molecular Biology Department,
Genentech, Inc. South San Francisco, California
1988 - present Senior Research Scientist, Molecular Oncology Department,
Genentech, Inc. South San Francisco, California
SOCIETIES
American Association for the Advancement of Science
International Cytokine Society
American Association for Cancer Research
AWARDS AND HONORS
1984-85
September 27, 1986
December 27, 1987
Chosen by Science Digest as One of the Scientists Involved in the Top 100
Innovations of the Year for Work on Human Tissue Plasminogen Activator
Distinguished Alumni Award, State University of New York, College at
Fredonia
Chosen by New York Times as One of Four People for the Article "The People
Behind Some of the Bright Ideas of 1987"
68
April 23, 1988
May 14, 1988-
April 13, 1989-
1990
June 30, 1991
November 1995
September 30, 1996
May, 1997
January 21, 1998
January 29, 1998
December, 1998
May 19, 1999
January 29, 2000
June, 2001
August 10,2001
July 15,2002
Service to Humanity Award, Fredonia Chamber of Commerce, Fredonia, New
York
Commencement Speech to College Graduates at State University of New York,
College at Fredonia
Inventor of the Year Award, Intellectual Property Owners Foundation
Genentech President s Award for Outstanding Achievement
Nominated for Induction into The National Inventors Hall of Fame
Named to the SUNY Alumni Honor Role for excellence in career achievements
(State University of New York)
Nominated for the Lemelson-MIT Prize for Inventors
Chosen to be included in a chapter in the book "Prescriptions for Profits:
How the Pharmaceutical Industry Bankrolled the Unholy Marriage Between
Science and Business" by Linda Marsa
Chosen by PhRMA to represent Genentech in television and print ads on the
impact and potential of t-PA for stroke patients
Chosen to speak to Vice president Al Gore when he visited Genentech to
announce two cancer-related proposals: an additional $4.7 billion a 65%
increase over five years in funding for cancer research at the National Institutes
of Health; and the expansion of Medicare benefits for cancer patients.
Chosen to be included in the book "Patently Female" under Medical
Innovations by Ethlie Ann Vare (General Publishing)
Invited Speaker at the Master of Liberal Studies in Technology Lecture Series
"21 st Century Inventing" Eastern Michigan University
One of three women profiled in a speech to the American Chemical Society by
author, Autumn Stanley on "Women Inventors Who Make a Difference"
Chosen to Give an Oral History on the t-PA project for the Bancroft Library
at the University of California at Berkeley for The Program in the History of the
Biological Sciences & Biotechnology
Invited Speaker for Career Symposium at Vanderbilt University to Graduate
Students and Post-Doctoral Fellows about Research in the Biotechnology Industry
Chosen as One of 31 Scientists Profiled on the Genentech Web Site out of 546
People in Research. See: http ://www. gene .coin/g ene/researc h/sc i -pro fi les/
69
PUBLICATIONS
1. Walsh, M.L, Pcnnica, D. and Cohen, P.S. The Distribution of Functional
Bacteriophage T4 mRNA on Polysomes. Arch. Biochem. Biophys. 173. 732-738
(1976).
2. Walker, A.C., Walsh, M.L., Pennica, D., Cohen, P.S. and Ennis, H.L.
Transcription-translation and Translation-messenger RNA Decay Coupling:
Separate Mechanisms for Different Messengers. Proc. Nat. Acad. Sci. USA 73,
1126-1130(1976).
Pennica, D. and Cohen, P.S. Regulation of Ribosome Function Following
Bacteriophage T 4 Infection. J. Mol. Biol. 122. 137-144 (1978).
4. Ennis, H.L., Pennica, D. and Hill, J.M. Synthesis of Macromolecules during
Microcyst Germination in the Cellular Slime Mold Polysphondylium pallidum.
Dev. Biol. 65, 25 1 -259 ( 1 978).
Pennica, D., Lynch, K.R., Cohen, P.S. and Ennis H.L. Decay of Vesicular
Stomatitis Virus mRN As In Vivo. Virology 94, 484-487 ( 1 979).
6. Lynch, K.R., Pennica, D., Ennis, H.L. and Cohen, P.S. Separation and
Purification of the mRN As for Vesicular Stomatitis Virus NS and M Proteins.
Virology 98, 25 1 -254 ( 1 979).
Pennica, D., Holloway, B.P., Heyward, J.T. and Obijeski, J.F. In Vitro
Translation of Rabies Virus messenger RN As. Virology 103. 517-521(1 980).
Lynch, K.R., Pennica, D., Ennis, H.L. and Cohen, P.S. Temporal Regulation of
the Rate of Vesicular Stomatitis Virus mRNA Translation during Infection of
Chinese Hamster Ovary Cells. Virology 108. 277-285 ( 1 98 1 ).
9. Pennica, D., Cohen, P.S. and Ennis, H.L. Mechanism of Vesicular Stomatitis
Virus mRNA Decay. Archives of Biochemistry and Biophysics 208. 403-408
(1981).
10. Heyneker, H., Holmes, W., Rey, M., Pennica, D., Shepard H.M., Seeburg, P.,
Hayflick, J., Ward, C. and Vehar, G Functional Expression of the Human
Urokinase Gene in E. coli. Proceedings of the Fourth International Symposium
on Genetics of Industrial Microorganisms, 214-221 (1982).
1 1 Gray, P. W., Leung, D. W., Pennica, D., Yelverton, E., Najarian, R., Simonsen,
C.C., Derynck, R., Sherwood, P.J., Wallace, D.M., Berger, S.L., Levinson, A.D.
and Goeddel, D.V. Expression of Human Immune Interferon cDNA in E. coli and
Monkey Cells. Nature 225, 503-508 (1982).
1 2. Simonsen, C.C., Shepard, H.M., Gray, P. W, Leung, D. W., Pennica, D.,
Yelverton, E., Derynck, R., Sherwood, P.J., Levinson, A.D. and Goeddel, D.V.
70
Plasmid-Directed Synthesis of Human Immune Interferon in E. coli and Monkey
Cells. "Interferons, Proceedings of the UCLA Symposia on Molecular Biology
Vol.25" 1-14(1982).
1 3. Pennica, D., Holmes, W.E., Kohr, W.J., Harkins, R.N., Vehar, G A., Ward, C.A.,
Bennett, W.F., Yelverton, E., Seeburg, P.H., Heyneker, H.L., Goeddel, D.V. and
Collen, D. Cloning and Expression of Human Tissue-type Plasminogen Activator
cDNA in E. coli. Nature 301, 214-221 (1982).
14. Derynck, R., Gray, P.W., Yelverton, E., Leung, D.W., Shepard, H.M., Lawn,
R.M., Ullrich, A., Najarian, R., Pennica, D., Hagie, F.E., Hitzeman, R.A.,
Sherwood, P.J., Levinson, A.D. and Goeddel, D.V. Synthesis of Human
Interferons and Analogs in Heterologous Cells, "From Gene to Protein:
Translation into Biotechnology." Edited by F. Ahmad et al.. Academic Press,
New York, 249-262(1982).
1 5. Pennica, D., Holmes, W.E., Kohr, W.J., Harkins, R.N., Vehar, GA., Ward, C.A.,
Yelverton, E., Seeburg, PH., Heyneker, H.L., Goeddel, D.V. and Collen, D.
Human Tissue-type Plasminogen Activator: Properties of the mRNA, its cDNA
and the Translated Protein, "Progress in Fibrinolysis," Vol. VI. Edited by J.F.
Davidson et al.. Churchill Livingston, New York, 269-273 (1983).
16. Collen, D., Stassen, J.M., Marafino, B.J., Jr., Builder, S., De Cock, F., Ogez, J.,
Tajiri, D., Pennica, D., Bennett, W.F., Salwa, J. and Hoyng, C.F. Biological
Properties of Human Tissue-type Plasminogen Activator Obtained by Expression
of Recombinant DNA in Mammalian Cells. The Journal of Pharmacology and
Experimental Therapeutics 231. 146-152 (1984).
1 7. Pennica, D., Goeddel, D.V, Hayflick, J.S., Reich, N.C., Anderson, C. W. and
Levine, A.J. The Amino Acid Sequence of Murine p53 Determined from a cDNA
Clone. Virology 134. 477-482 (1984).
1 8. Vehar, GA., Kohr, W.J., Bennett, W.F., Pennica, D., Ward, C.A., Harkins, R.N.
and Collen, D. Characterization Studies on Human Melanoma Cell Tissue
Plasminogen Activator. Bio/Technology 2, 1051-1057 (1984).
19. Pennica, D., Nedwin, GE., Hayflick, J.S., Seeburg, PH., Derynck, R., Palladino,
M.A., Kohr, W.J., Aggarwal, B.B. and Goeddel, D.V. Human Tumour Necrosis
Factor: Precursor Structure, Expression, and Homology to Lymphotoxin. Nature
312.724-729(1984).
20. Pennica, D., Hayflick, J.S., Palladino, M.A. and Goeddel, D.V. Cloning and
Expression in Escherichia coli of the cDNA for Murine Tumor Necrosis Factor.
Proc. Nat. Acad. Sci. USA 82, 6060-6064 (1985).
21. Nedwin, GE., Naylor, S.L., Sakaguchi, A.Y., Smith, D., Jarrett-Nedwin, J.,
Pennica, D., Goeddel, D.V. and Gray, P.W. Human Lymphotoxin and Tumor
Necrosis Factor Genes: Structure, Homology and Chromosomal Localization.
Nucleic Acids Research U, 6361-6373 (1985).
71
Holmes, W.E., Pennica, D., Blaber, M., Key, M.W., Guenzler, W.A., Steffens,
QJ. and Heyneker, H.L. Cloning and Expression of the (iene for Pro-Urokinase
in Escherichia coli. Bio/Technology 3_, 923-929 ( 1 985).
23. Aggarwal, B.B., Hass, P.E., Kohr, W.J., Nedwin, GE., Gray, P.W., Pennica, I).,
Svedersky, L.P., Lee, S.H., Palladino, M.A. and Goeddel, D.V. Human Tumor
Necrosis Factors Alpha and Beta: Their Biochemical and Biological Properties.
In Recent Advances in Chemotherapy: Anticancer Section. Proceedings of the
14th International Congress of Chemotherapy - Kyoto. Edited by J. Ishigami.
Univesity of Tokyo Press, Tokyo, 83-85 (1985).
24. Shalaby, M.R., Pennica, D. and Palladino, M.A., Jr. An Overview of the History
and Biologic Properties of Tumor Necrosis Factors. Springer Seminars in
Immunopathology 9, 33-37 (1986).
25. Gray, P.W., Glaister, D., Chen, E., Goeddel, D.V. and Pennica, D. Two
Interleukin 1 Genes in the Mouse: Cloning and Expression of the cDNA for
Murine Interleukin Ib. J. Immunol. 137. 3644-3648 (1986).
26. Goeddel, D.V., Aggarwal, B.B., Gray, P.W., Leung, D.W., Nedwin, GE.,
Palladino, M.A., Patton, J.S., Pennica, D., Shepard, H.M., Sugarman, B.J. and
Wong, GH.W. Tumor Necrosis Factors: Gene Structure and Biological
Activities. In Cold Spring Harbor Symposium on Quantitative Biology, Vol. 51,
597-609(1986).
27. Pennica, D. and Goeddel, D.V. Cloning and Characterization of the Genes for
Human and Murine Tumor Necrosis Factors. In Lymphokines: Molecular
Cloning and Analysis of Lymphokines, Vol. 13, Academic Press, 163-180 (1987).
28. Pennica, D., Shalaby M.R. and Palladino, M.A., Jr. Tumor Necrosis Factors
Alpha and Beta. In Recombinant Lymphokines and Their Receptors. Edited by
Steven Gillis. Marcel Dekker, Inc., p. 301-317, (1987).
29. Pennica, D., Kohr, W.J., Kuang, W.J., Glaister, D., Aggarwal, B.B., Chen, E.Y.
and Goeddel, D.V. Identification of Human Uromodulin as the Tamm-Horsfall
Urinary Glycoprotein. Science 236. 83-88 (1987).
30. Aggarwal, B.B., Aiyer, R.A., Pennica, D., Gray, P.W. and Goeddel, D.V. Human
Tumor Necrosis Factors: Structure and Receptor Interactions. Edited by Joan
Marsh and Julie Whelan. CIBA Foundation Symposium, No. 131, on Tumor
Necrosis Factors and Related Cytotoxins, 39-51 (1987).
31. Rice, GC. and Pennica, D. Detection by Flow Cytometry of Protoplast Fusion
and Transient Expression of Transferred Heterologous CD4 Sequences in COS-7
Cells. Cytometry JO, 103-107(1989).
32. Rice, GC., McCabe, S., Pennica, D., Woronicz, J., Borree, J. and Goeddel, D.V.
Expression Cloning by Transient Expression Utilizing Fluorescence Activated
Cell Sorting. DNA & Protein Engineering Techniques 2, 1 8-23 (1990).
72
33. Rice, G.C., Pennica, D., Borree, J.A. and Williams, S.R. Measurement of
Transient cDNA Expression in Mammalian Cells Using Flow Cytometric Cell
Analysis and Sorting. Cytometry JL2, 22 1 -223 ( 1 99 1 ).
34. Gibbs, V.C., Williams, S.R., Gray, P.W., Schreiber, R.D., Pennica, D., Rice, G
and Goeddel, D.V. The Extracellular Domain of the Human Interferon Gamma
Receptor Interacts with a Species-Specific Signal Transducer. Mol. Cell Biol. JJL,
5860-5866(1991).
35. Ashkenazi, A., Marsters, S.A., Capon, D.J., Chamow, S.M., Figari, I.S., Pennica,
D., Goeddel, D.V., Palladino, M.A. and Smith, D.H. Protection Against
Endotoxic Shock by A Tumor Necrosis Factor Immunoadhesin. Proc. Nat l.
Acad. Sci. USA 88, 10535-10539 (1991).
36. Pennica, D., Kohr, W.J., Fendly, B.M., Shire, S.J., Raab, H.E., Borchardt, P.E.,
Lewis, M. and Goeddel, D.V. Characterization of a Recombinant Extracellular
Domain of the Type 1 TNF Receptor: Evidence for TNF-alpha Induced Receptor
Aggregation. Biochemistry 31, 1134-1141 (1992).
37. Pennica, D., Lam, V.T., Mize, N.K., Weber, R.F., Lewis, M., Fendly, B.M.,
Lipari, M.T. and Goeddel, D.V. Biochemical Properties of the 75kDa Tumor
Necrosis Fator Receptor. J. Biol. Chemistry 267. 2 1 1 7 1 -2 1 1 78 ( 1 992).
38. Pennica, D., Lam, V.T., Weber, R.F., Kohr, W.J., Basa, L.J., Spellman, M.W.,
Ashkenzai, A., Shire, S.J. and Goeddel, D.V. Biochemical Characterization of the
Extracellular Domain of the 75 kDa Tumor Necrosis Factor Receptor.
Biochemistry 32, 3131-3138 (1993).
39. Greenlund, A.C., Schreiber, R.D., Goeddel, D.V., and Pennica, D. Interferon-y
Induces Receptor Dimerization in Solution and on Cells. J. Biol. Chem. 268.
18103-18110(1993).
40. Tartaglia, L.A., Pennica, D., and Goeddel, D.V. Ligand Passing: The 75-kDa
Tumor Necrosis Factor (TNF) Receptor Recruits TNF for Signaling by the 55-
kDa TNF Receptor. J. Biol. Chem. 268. 18542-18548 (1993)
41 . Pennica, D., King, K. L., Shaw, K. J., Luis, E., Rullamus, J., Luoh, S-M.,
Darbonne, W. C., Knutzon, D. S., Yen, R., Chien, K. R., Baker, J. B., and
Wood,W. 1. Expression Cloning of Cardiotrophin- 1 , a Cytokine that Induces
Cardiac Myocyte Hypertrophy Proc. Nat l. Acad. Sci. USA 92, 1 142-1 146
(1995).
42. Marsters. S..A., Pennica, D., Bach, E., Schreiber, R.D., and Ashkenazi, A.
Interferon g signals via a high-affinity multisubunit receptor complex that
contains two types of polypeptide chain. Proc. Nat l. Acad. Sci. USA 92. 5401-
5405(1995).
43. Pennica, D., Shaw, K. J., Luoh, S-M., and Wood, W. I. Isolation of cDNA Clones
Encoding the Mouse Protein V-l. Gene, 158. 305-306, (1995).
73
44. Pennica, D., Shaw, K. J., Swanson, T. A., Moore, M. W., Shclton, D. L.,
Zioncheck K. A., Roscnthal. A., Taga, T., Paoni, N. F., and Wood. W. I.
Cardiotrophin-1: Biological Activities and Binding to the Leukemia Inhibitory
Factor Receptor/gpl30 Signaling Complex. J. Biol. Chem. 270. 10915-10922,
(1995).
45. Peters, M., Roeb, E., Pennica, D., BUschenfelde, K-H. M., and Rose-John, S. A
New Hepatocyte Stimulating Factor: Cardiotrophin-l (CT-I), FEBS Lett. 372:
177- 1 80, (1995)
46. Habecker, B.A., Pennica, D., and Landis, S. C., Cardiotrophin-1 is not the Sweat
Gland Differentiation Factor Neuroreport, 7, 41-44, (1995)
47. Richards, C. D., Langdon, C., Pennica, D., and Gauldie, J., Murine
Cardiotrophin-1 Stimulates the Acute Phase Response in Rat Hepatocytes and
H35 Hepatoma Cells Journal of Interferon and Cytokine Research, 16. 69-75,
(1996)
48. Benigni, F., Fantuzzi, G, Sacco, S., Sironi, M., Pozzi, P., Dinarello, C. A., Sipe, J.
D., Poll, V., Cappelletti, M., Paonessa, G, Pennica, D., Panayotatos, N., and
Ghezzi, P. Six Different Cytokines that Share gp!30 as a Receptor Subunit,
Induce Serum Amyloid A and Potentiate the Induction of Interleukin-6 and the
Activation of the Hypothalamus-Pituitary-Adrenal Axis by Interleukin-1 Blood,
S7, 1851-1854, (1996)
49. Pennica, D., Swanson, T. A., Shaw, K. J., Kuang, W-J., Gray, C., Beatty, B. G,
and Wood. W. 1. Human Cardiotrophin-1: Protein and Gene Structure, Biological
and Binding Activities and Chromosomal Localization. Cytokine 8, 183-189,
(1996)
50. Sheng, Z., Pennica, D., Wood, W.I., and Chien, K. R., Cardiotrophin-1 Displays
Early Expression in the Murine Heart Tube and Promotes Cardiac Myocyte
Survival. Development, 122. 4 1 9-428, ( 1 996)
5 1 . Wollert, K. C., Taga, T., Saito, M., Narazaki, M., Kishimoto, T, Glembotski,
C.C., Vernallis, A. B., Heath, J. K., Pennica, D., Wood, W. I., and Chien, K. R.
Cardiotrophin-1: Activates a Distinct Form of Cardiac Muscle Cell Hypertrophy:
Assembly of Sarcomeric Units in Series via gp!30/Leukemia Inhibitory Factor
Receptor-Dependent Pathways J. Biol. Chem. 271. 9535-9545, (1996)
Pennica, D., Wood, W. I., and Chien, K. R., Cardiotrophin- 1 : A Multifunctional
Cytokine that Signals via L1F Receptor-gpl30 Dependent Pathways, Cytokine
and Growth Factor Reviews, 7, 8 1 -9 1 ,( 1 996)
Benigni, F., Sacco, S., Pennica, D., and Ghezzi, P., Cardiotrophin-1 Inhibits TNF
Production in the Heart and Serum of Lipopolysaccharide-Treated Mice and in
vitro in Mouse Blood Cells, American Journal of Pathology 149, 1847-1850
(1996)
74
54. Jin, H., Yang, R., Keller, G, Ryan, A., Ko, A., Finkle, D., Swanson, T. A., Li, W.,
Pennica, D., Wood, W.I., and Paoni, N.F., In Vivo Effects of Cardiotrophin-1
Cytokine, 8, 920-926 (1996)
55. Pennica, D., Arce, V., Swanson, T. A., Vejsada, R., Pollock, R. A., Armanini,
M., Dudley, K., Phillips, H. S., Rosenthal, A., Kato, A., and Henderson, C. E.,
Cardiotrophin-1, A Cytokine Present in Embryonic Muscle, Supports Long Term
Survival of Spinal Motorneurons Neuron, 17, 63-74 (1996)
56. Stone, D., Hynes, M., Armanini, M., Swanson, T. A., Gu, Q., Johnson, R. L.,
Scott, M. P., Pennica, D., Goddard, A., Phillips, H., Noll, M., Hopper, J. E.,
deSauvage, F., and Rosenthal, A. The Tumour-Supressor Gene Patched Encodes
a Candidate Receptor for Sonic Hedgehog Nature, 384, 129-134 (1996)
57. Gibbs, V. C., and Pennica, D., CRF2-4: Isolation of cDNA Clones Encoding the
Human and Mouse Proteins Gene 186, 97-101 (1997)
58 Robledo, O., Fourcin, M., Chevalier, S., Guillet, C., Auguste, P., Pouplard-
Barthelaix, A., Pennica, D., and Gascan, H., Signalling of the Cardiotrophin-1
Receptor : Evidence for a Third Receptor Component J. Biol. Chem., 272, 4855-
4863(1997)
59. Cheng, J G, Pennica, D., and Patterson, P. H. Cardiotrophin-1 Induces the
Same Neuropeptides in Sympathetic Neurons as do Neuropoietic Cytokines, J.
Neurochem., 69, 2278-2284 (1997)
60. Robledo, O., Guillet, C., Chevalier, S., Fourcin, M., Froger, J., Pouplard-
Barthelaix, A., Pennica, D., and Gascan, H. Hepatocyte Derived Cell Lines
Express a Functional Receptor for Cardiotrophin-1. Eur.Cytokine Netw., 8, 245
252(1997).
61 . Robledo, O., Chevalier, S., Froger, J., Barthelaix-Pouplard, A., Pennica, D., and
Gascan, H. Regulation of Interleukin-6 Expression by Cardiotrophin-1 Cytokine
9,666-671(1997)
62. Richards, C. D., Kerr, C., Tanaka, M., Hara, T., Miyajima, A., Pennica, D.,
Botelho, F., and Langdon, C. M. Regulation of Tissue Inhibitor of
Metalloproteinase- 1 in Fibroblasts and Acute Phase Proteins in Hepatocytes In
Vitro by Mouse Oncostatin M, Cardiotrophin-1 and Interleukin-6 Journal of
Immunology, 159, 2431-2437 (1997)
63. Pennica, D., and Wood, W.I., Cardiotrophin-1 in Human Cytokines: Handbook
for Basic and Clinical Research, Volume III Blackwell Science, Inc. 1-16 (1998)
64. Jin, H., Yang, R., Ko, A., Pennica, D., Wood, W. I., and Paoni, N. F. Effects of
Cardiotrophin-1 on Hemodynamics and Cardiac Function in Conscious Rats
Cytokine, 10, 19-25(1998)
65. Stephanou, A., Brar, B., Heads, R., Knight, R.D., Marber, M.S., Pennica, D., and
Latchman, D.S., Cardiotrophin-1 Induces Heat Shock Protein Accumulation in
75
Cultured Cardiac Cells and Protects Them from Stressful Stimuli J. Mol. Cell.
Cardiology, 30 (4), 849-855, (1998)
66. Chandrasekar, B., Melby, C. P., Pennica, D., and Freeman, G L.
Overexpression of Cardiotrophin-1 and gp!30 during Experimental Acute
Chagasic Cardiomyopathy Immunol. Lett., 61(2-3), 89-95 (1998)
67. Morton, A. R., Bartlett, P. F., Pennica, D., Davies, A. M., Cytokines Promote the
Survival of Cranial Sensory Neurons at Different Developmental Stages, Eur. J.
Neuroscience, 10, 673-679, (1998)
68. Arce, V., Pollock, R. A., Philippe, J.M., Pennica, D., Henderson, C. E., and
deLapeyriere, O., Snergistic Effiects of Schwann- and Muscle-Derived Factors on
Motoneuron Survival Involve GDNF and CT-I, J. Neuroscience, 18(4), 1440-
1448, (1998)
69. Geissen, M., Heller, S., Pennica, D., Emsberger, U., and Rohrer, H., The
Specification of Sympathetic Neurotransmitter Phenotype Depends on gp!30
Cytokine Receptor Signaling, Development, 125, 4791-4801 (1998)
70. Rodig, S. J., Meraz, M. A., White, H. M., Lampe, P., Riley, J., Arthur, C., King,
K.., Sheehan, K. C. F., Pennica, D., Johnson, E., and Schreiber, R. D., Targeted
Disruption of the Jakl Gene Reveals the Obligatory and Nonredundant Roles of
Janus Kinases in Mediating Cytokine Induced Biologic Responses, Cell, 93. 373-
383(1998)
7 1 . Pennica, D., Swanson, T. A., Welsh, J. W., Roy, M. A., Lawrence, D. A., Lee, J.,
Brush, J. ,.Taneyhill, L. A., Deuel, B., Lew, M., Watanabe, C., Cohen, R. L.,
Melham, M. F., Finley, G G, Quirke, P., Goddard, A. D., Hillan, K. J. Gumey, A.
L., Botstein, D., and Levine, A.J., WISP Genes are Members of the Connective
Tissue Growth Factor Family that are Up-Regulated in Wnt-1 Transformed Cells
and Aberrantly Expressed in Human Colon Tumors Proc. Natl. Acad. Sci., 95,
14717-14722(1998)
72. Aoyama, T., Takimoto, Y., Pennica, D., Inoue, R., Shinoda, E., Hattori, R., Yui,
Y., Sasayama, S., Augmented Gene Expression of Cardiotrophin- 1 and Its
Receptor Component , gp!30, in Both Left and Right Ventricles after Myocardial
Infarction in the Rat Circulation (suppl) 98:839,(1998).
73. Pulido, E.J., Shames, B. D., Pennica, D., O Leary, R. M., Bensard, D. D., Cain,
B. S., Mclntyre, R. C. Cardiotrophin-1 Attenuates Endotoxin-Induced Acute Lung
Injury J. Surg. Res, 84, 240-246 ( 1 999)
74. Thier, M., Hall, M., Heath, J.K., Pennica, D., and Weis, J., Trophic Effects of
Cardiotrophin-1 and Interleukin-1 1 on Newborn Rat Sensory Dorsal Root
Ganglion Neurons in Vitro Molecular Brain Research, 64, 80-84 (1999)
Richards, C.D., Langdon, C., Deschamps, P., Pennica, D., and Shaughnessy, S.
G, Stimulation of Osteoclast Differentiation in vitro by Mouse Oncostatin M,
76
Leukemia Inhibitory Factor, Cardiotrophin- 1 and Interleukin-6: Synergy with
Dexamethazone Cytokine 12, No.6 613-621(2000)
76. Middleton, G, Hamanoue, M., Enokido, Y, Wyatt, E. J., Pennica, D., Hay, R. T,
and Davies, A. M. Cytokine Induced NF-kB Activation Promotes the Survival of
Developing Neurons , J. Cell Biol. Jan 24;148(2):325-32 (2000)
77. Xu, L., Corcoran, R., Welsh, J., Pennica, D., and Levine, A.J. WISP-1 is a Wnt-
land b-catenin Responsive Oncogene Genes & Development, 14, 585-595 (2000)
78. Fryer, H.J.., Wolf, D. H., Knox, R. J., Strittmatter, S. M., Pennica, D., O Leary,
R. M., Russell, D., and Kalb, R. G, Brain Derived Neurotrophic Factor Induces
Excitoxic Sensitivity in Cultured Embryonic Rat Spinal Motor Neurons Through
Activation of the Phosphatidylinositol 3-K.inase Pathway J. Neurochem.
Feb;74(2):582-95 (2000)
79. Railson, J., Lawrence, K.., Stephanou, A., Brar, B. K., Pennica, D., and
Latchman, D. S. Cardiotrophin- 1 (CT-1) Reduces Stress Induced Heat Shock
Protein Production in Cardiac Myocytes Cytokine, 12, 1741-1744, (2000)
80. Aoyama, T., Takimoto, Y., Pennica, D., Inoue, R., Shinoda, E., Hattori, R., Yui,
Y, Sasayama, S., Augmented Expression of Cardiotrophin- 1 and Its Receptor
Component , gp!30, in Both Left and Right Ventricles after Myocardial Infarction
in the Rat J Mol Cell Cardiol, 32 1821-1830, (2000).
81. Taneyhill, L. A., Pennica, D., and Levine, A. J. Identification of a Mouse
Homolog of the Human BTEB2 Transcription Factor as a b-Catenin-Independent
Wnt-1 Response Gene, Mol. Cell. Biol. 21: 562-574 (2001)
82. Oppenheim, R. W., Wiese, S., Prevette, D., Armanini, M., Wang, S., Houenou,
L.J., Holtmann, B., Gotz, R., Pennica, D., and Sendtner, M., Cardiotrophin- 1
(CT-1), a Muscle-Derived Cytokine, is Required for the Survival of
Subpopulations of Developing Motoneurons, J Neurosci. Feb 15; 2 1(4): 1283-
1291 (2001)
83. Szeto, W., Jiang, W., Tice, D.A., Rubinfeld, B., Hollingshead, P.G, Fong, S.E.,
Dugger, D. L., Pham, T., Yansura, D. G, Wong, T. A., Grimaldi, J.C., Corpuz, R.
T., Singh, J. S., Frantz, G D., Devaux, B., Crowley, C.W., Schwall, R. H.,
Eberhard, D. A., Rastelli, L., Polakis, P., and Pennica, D., Overexpression of the
Retinoic Acid-Responsive Gene Stra6 in Human Cancers and its Synergistic
Induction by Wnt-1 and Retinoic Acid Cancer Research, May 15; 61 4197-4205
(2001)
84. Mitsumoto H, Klinkosz B, Pioro EP, Tsuzaka K, Ishiyama T, O Leary RM,
Pennica, D., Effects of Cardiotrophin- 1 (CT-1) in a mouse motor neuron disease.
Muscle Nerve. Jun;24(6):769-77, (2001)
n
Ayer-Lelievre C, Brigstock D, Lau L, Pennica I). Perbal B. Ycger M. Report and
abstracts on the first international workshop on the C C N family of genes. Mol
Pathol. Apr; 54(2): 105-20. (2001)
86. Railson, J. E., Lawrence, K., Buddie, J. C., Pennica, D., and Latchman, D. S.
Heat Shock Protein-56 is Induced by Cardiotrophin- 1 and Mediates its
Hypertrophic Effect , J Mol Cell Cardiol. Jun;33(6): 1209-21 (2001)
87. Tao, W., Pennica, D., Xu, L., Kalejta, R. F., and Levine, A. J., Wrch-1, a novel
member of the Rho gene family that is regulated by Wnt-1 Genes &
Development, Vol. 15, No. 14, pp. 1796-1807, July 15, (2001)
88. Brar, B.K., Stephanou, A., Liao, Z., O Leary, R. M., Pennica, D., Yellon, D.M.,
and Latchman, D. S Cardiotrophin- 1 Can protect Cardiac Myocytes from Injury
When Added Both Prior to Simulated Ischaemia and at Reoxygenation
Cardiovascular Research, Aug l;51(2):265-74) (2001)
89. Desnoyer, L., Arnott, D., and Pennica, D., W1SP-1 Binds to Decorin and
Biglycan Journal of Biological Chemistry, 276, No. 50 47599-47607 (2001)
90. Brar, B.K., Stephanou, A., Pennica, D., and Latchman, D. CT-1 Mediated
Cardioprotection Against Ischaemic Re-Oxygenation Injury Is Mediated By Pi3
Kinase, Akt And Mekl/2 Pathways Cytokine, Nov; 16(3):93-6 (2001)
91. Takimoto Y, Aoyama T, Iwanaga Y, Izumi T, Kihara Y, Pennica D, Sasayama S.
Increased expression of Cardiotrophin- 1 during ventricular remodeling in
hypertensive rats. Am J Physiol Heart Circ Physiol Mar; 282(3):H896-90 1(2002)
92. Liao, Z, Brar, BK, Cai, Q, Stephanou, A, O Leary RM, Pennica, D, Yellon, D.M.,
Latchman D.S. Cardiotrophin- 1 (CT-1 ) Can Protect the Heart From Injury When
Added Both Prior to Ischaemia and at Reperfusion Cardiovascular Research
Mar;53(4):902-l 0(2002)
93. Railson, J.E., Liao, Z., Brar, B.K., Pennica, D., Stephanou, A., and Latchman,
D.S. Cardiotrophin- 1 and Urocortin Cause Protection by the Same Pathway and
Hypertrophy via Distinct Pathways in Cardiac Myocytes Cytokine, Mar
7;17(5):243-53(2002)
94. Tice, D. A., Szeto, W., Soloviev, L, Rubinfeld, B., Fong, S.E., Dugger, D.L.,
Winer, J., Williams, P. M., Wieand, D., Smith, V., Schwall, R. H., Pennica, D.,
and Polakis, P., "Synergistic Induction Of Tumor Antigens By Wnt-1 Signaling
And Retinoic Acid Revealed By Gene Expression Profiling J Biol Chem. Apr 19;
277(1 6): 14329-35 (2002)
95. Gajavelli, S., Wood, P., Pennica, D., Whittemore, S., and Pantelis, Tsoulfas BMP
signaling initiates a neural crest differentiation program in embryonic rat CNS
stem cells. Exp Neurol. Aug;188(2):205-23 (2004)
78
96. Gard, A. L., Gavin, E., Solodushko, V, Pennica, D. Cardiotrophin-1 in Choroid
Plexus and the Cerebrospinal Fluid Circulatory System,
Neuroscience.; 1 27( 1 ):43-52. (2004)
97. Taneyhill, L., and Pennica, D., Identification of Wnt Responsive Genes Using a
Murine Mammary Epithelial Cell Line Model System, BMC Developmental
Biology 4:6 1-14(2004)
98. Sendtner, M., ... and Pennica, D. "CNTF, CT-1 and LIF - Triple-Knockout Mice.
" (in preparation) (2004)
99. Desnoyer, L., ... Pennica, D., and French, D. VV1SP-1 Promotes Metastasis by
Induction of Hyaluronan and CD44 (in preparation) (Nature Medicine) (2004)
PATENTS
WO0053758
Compositions and methods for the treatment of immune related diseases
Ashkenazi, Avi, J.; Baker, Kevin, P.; Goddard, Audrey; Gurney, Austin, L.; Hebert, Caroline; Henzel,
William; Kabakoff, Rhona, C.; Lu, Yanmei; Pan, James; Pennica, Diane; Shelton, David, L.; Smith,
Victoria; Stewart, Timothy, A.; Tumas, Daniel; Watanabe, Colin, K.; Wood, William, I.; Yan, Minhong
Application No. US0005841, Filed 20000302, A2 Published 20000914
WO0043790
Compositions and methods for the treatment of tumor
Botstein, David; Goddard, Audrey; Lawrence, David, A.; Pennica, Diane; Roy, Margaret, Ann; Wood,
William,
Application No. US0001441, Filed 200001 19, A2 Published 20000727
WOOO 15796
Secreted and transmembrane polypeptides and nucleic acids encoding the same
Chen, Jian; Goddard, Audrey; Gumey, Austin, L. Hillan, Kenneth; Pennica, Diane; Wood, William; Yuan,
Jean
Application No. US9921090, Filed 19990915, A2 Published 20000323
WO9921999
WISP Polypeptides and nucleic acids encoding the same
lnventor(s): Levine, Arnold, J.; Pennica, Diane
Application No. US9822992, Filed 19981029, A2 Published 19990506
W09921998
Wnt-1 induced secreted polypeptides: WISP-1, -2 and -3
Botstein, David, A.; Cohen, Robert, L.; Gumey, Austin, L.; Hillan, Kenneth; Lawrence, David, A.; Levine,
Arnold, J.; Pennica, Diane; Roy, Margaret, Ann; Goddard, Audrey; Wood, William, I.
Application No. US9822991, Filed 19981029, A 1 Published 19990506
79
WOW 14328
Secreted and transmembrane polypeplides and nucleic acids encoding the same
Wood, William, I.; Gumey, Austin, L.; Goddard, Audrey; Pennica, Diane; Chen, Jian; Yuan, Jean
Application No. US98 19330, Filed 19980916, A2 Published 19990325
WO9529237
Cardiotrophin and uses therefor
Inventors): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane ; Wood, William
Application No. US9504467, Filed 19950406, A 1 Published 19951 102
W09730146
Cardiotrophin and uses therefor
Inventors): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. US9702675, Filed 1997021 1, A2 Published 19970821
US5869314
Tissue plasminogen activators and derivatives thereof as produced by recombinant means
Inventors): Goeddel, David V.; Kohr, William J.; Pennica, Diane; Vehar, Gordon A.
Application No. 487456, Filed 19950606, Issued 19990209
US4766075
Human tissue plasminogen activator
Inventor(s): Goeddel, David V.; Kohr, William J.; Pennica, Diane ; Vehar, Gordon A.
Application No. 483052, Filed 19830407, Issued 19880823
US4853330
Human tissue plasminogen activator
Inventor(s): Goeddel, David V.; Kohr, William J.; Pennica, Diane ; Vehar, Gordon A.
Application No. 184477, Filed 19880421, Issued 19890801
US5011795
Human tPA production using vectors coding for DHFR protein
Inventors): Levinson, Arthur D.; Pennica, Diane; Kohr, William J.; Vehar, Gordon A.; Goeddel, David
V; Yelverton, Elizabeth M.; Simonsen, Christian C.
Application No. 499201, Filed 19900322, Issued 19910430
US5 185259
Truncated human tissue plasminogen activator
Inventor(s): Goeddel, David V; Kohr, William J.; Pennica, Diane; Vehar, Gordon A.
Application No. 489855, Filed 19900302, Issued 19930209
US5268291
Human t-PA production using vectors coding for DHFR protein
Inventor(s): Levinson, Arthur D.; Pennica, Diane; Kohr, William J.; Vehar, Gordon A.; Goeddel, David V.
; Simonsen, Christian C.
Application No. 663103, Filed 19910228, Issued 19931207
US5424198
liuman t-PA production using vectors coding for DHFR protein
Inventors): Levinson, Arthur D.; Pennica, Diane; Kohr, William J.; Vehar, Gordon A.; Goeddel, David
V.; Yelverton, Elizabeth M.; Simonsen, Christian C.
Application No. 162354, Filed 19931203, Issued 19950613
US5571893
Cardiac hypertrophy factor
Inventors): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 286304, Filed 19940805, Issued 19961105
US5571675
Detection and amplification of candiotrophin-1 (cardiac hypertrophy factor)
Inventors): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 444083, Filed 19950517, Issued 19961105
US5587159
Human tissue plasminogen activator
Inventor(s): Goeddel, David V.; Kohr, William J.; Pennica, Diane; Vehar, Gordon A.
Application No. 264134, Filed 19940621, Issued 19961224
US5624806
Antibodies to cardiac hypertrophy factor and uses thereof
Inventor(s): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 442745, Filed 19950517, Issued 19970429
US5627073
Hybridomas producing antibodies to cardiac hypertrophy factor
Inventor(s): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 443129, Filed 19950517, Issued 19970506
US5849574
Human t-PA production using vectors coding for DHFR protein
Inventors): Levinson, Arthur D.; Pennica, Diane; Kohr, William J.; Vehar, Gordon A.; Goeddel, David
V.; Yelverton, Elizabeth M.;Simonsen, Christian C.
Application No. 450874, Filed 19950526, Issued 19981215
US5702938
Human tissue plasminogen activator
Inventor(s): Goeddel, David V; Kohr, William J.; Pennica, Diane; Vehar, Gordon A.
Application No. 468974, Filed 19950606, Issued 19971230
US5679545
Gene encoding cardiac hypertrophy factor
Inventors): Baker, Joffre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 443952, Filed 19950517, Issued 19971021
81
US5723585
Method of purifying cardiac hypertrophy factor
Inventor(s): Baker, Jofl rc; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. 443130, Filed 19950517, Issued 19980303
US5728566
Tissue plasminogen activator derivatives
Inventor(s): Goeddel, David V.; Kohr, William J. ; Pennica, Diane ; Vehar, Gordon A.
Application No. 483571, Filed 19950606, Issued 19980317
US5728565
Methods of preparing tissue plasminogen activator derivatives
Inventor(s): Goeddel, David V. Kohr, William J. ; Pennica, Diane ; Vehar, Gordon A.
Application No. 210179, Filed 19940317, Issued 19980317
US5753486
Human tissue plasminogen activator
Inventor(s): Goeddel, David V.; Kohr, William J. ; Pennica, Diane ; Vehar, Gordon A.
Application No. 472549, Filed 19950606, Issued 19980519
US5763253
Methods of preparing tissue plasiminogen activator derivative composition
Inventors): Goeddel, David V. ; Kohr, William J. ; Pennica, Diane ; Vehar, Gordon A.
Application No. 474160, Filed 19950606, Issued 19980609
JP020 16981
DNA coding for human tissue plasminogen activation factor
Inventors): Goeddel David V Kohr William J ; Pennica Diane ; Vehar Gordon A
Application No. 01023654, Filed 19890201, Published 19900119
JP05 137583
Vector containing DNA encoding human tissue plasminogen activator
Inventors): Goeddel, David V, Kohr, William J; Pennica, Diane; Vehar Gordon A
Application No. 04096448, Filed 19920416, Published 19930601
EP 1029046
Wnt-1 inducible genes
Inventors): Levine, Arnold, J. ;Pennica, Diane
Application No. EP98954038, Filed 19981029, A2 Published 20000823
U.S. Patent Number 6,387,657. U.S.
EP1027437
Wnt-1 induced secreted polypeptides: WISP-1, -2 and -3
Botstein, David, A.;Cohen, Robert, L.;Gumey, Austin, L.; Hillan, Kenneth; Lawrence, David, A.; Levine,
Arnold, J.; Pennica, Diane ;Roy, Margaret, Ann ; Goddard, Audrey ;Wood, William, I.
Application No. EP98956340, Filed 19981029, Al Published 20000816
82
EP 1027434
Secreted and transmembrane polypeptides and nucleic acids encoding the same
Wood, William, I. ; Gumey, Austin, L. ; Goddard, Audrey ; Pennica, Diane ; Chen, Jian ; Yuan, Jean
Application No. EP98946090, Filed 19980916, A2 Published 20000816
EP0755446
Cardiotrophin and uses therefor
Inventors): Baker, JofTre; Chien, Kenneth; King, Kathleen; Pennica, Diane; Wood, William
Application No. EP95921220, Filed 19950406, A 1 Published 19970129
EP0885294
Cardiotrophin and uses therefor
Inventor(s):Baker, Joffre ; Chien, Kenneth ; King, Kathleen; Pennica, Diane; Wood, William
Application No. EP97907730, Filed 19970211, A2 Published 19981223
PCT/USO 1/2 1635.
Methods for enhancing the efficacy of cancer therapy.
Inventors): Tice, D.A., Szeto, W., Pennica, D., Polakis, P.
US6472585
Cardiotrophin- 1 defective mouse
Inventors: Botstein; David; Goddard; Audrey Lawrence; David A.; Pennica; Diane Roy; Margaret Ann
Wood; William I.
Application No. 648183 Filed: August 25, 2000 Published October 29, 2002
US6387657
WISP polypeptides and nucleic acids encoding same
Inventors: Botstein; David A.; Cohen; Robert L.; Goddard; Audrey D.; Gurney; Austin L. ; Hillan;
Kenneth J.; Lawrence; David A.; Levine; Arnold J.; Pennica; Diane; Roy; Margaret Ann ; Wood; William
I.
Application No. 182145 Filed: October 29, 1998 Published: May 14, 2002
US6284247
Human tissue plasminogen activators
Inventors: Goeddel; David V.; Kohr; William J.; Pennica; Diane ; Vehar; Gordon A.
Application No. 105681 Filed: June 26, 1998 Published: September 4, 2001
US6274335
Method of treatment using recombinant human tissue plasminogen activator
Inventors: Goeddel; David V.; Kohr; William J.; Pennica; Diane; Vehar; Gordon A.
Application No. 105698 Filed: June 26, 1998 Published: August 14,2001
US6261837
Human tPA production using vectors coding for DHFR protein
Inventors: Levinson; Arthur D.; Pennica; Diane; Kohr; William J. ; Vehar; Gordon A.; Goeddel; David V;
Yelverton; Elizabeth M.; Simonsen; Christian C.
Application No. 105412 Filed: June 26, 1998 Published: July 17, 2001
83
US4853330
Human tissue plasminogen activator
Inventors: Goeddel; David V.; Kohr; William J.; Pennica; Diane; Vehar; Gordon A.
Application No. 184477 Filed: April 21, 1988 Published: August I, 1989
US20020 146707
Cardiotrophin-1 compositions and methods for the treatment of tumor
Inventors: Botstein, David;; Goddard, Audrey; Lawrence, David A.; Pennica, Diane; Roy, Margaret Ann;
Wood, William 1.;
Application No. 901257 Filed: July 9, 2001 Published: October 10, 2002
US 20020 137 189
Cardiac hypertrophy factor and uses therefore
Inventors: Baker, Joffre; Chien, Kenneth;; King, Kathleen; Pennica, Diane;; Wood, William;
Application No. 896856 Filed: June 29, 2001 Published: September 26, 2002
US20020 102622
Cardiotrophin- 1 compositions and methods for the treatment of tumor
Inventors: Botstein, David; Goddard, Audrey; Lawrence, David A.; Pennica, Diane; Roy, Margaret Ann;
Wood, William 1.;
Application No. 901540 Filed: July 9, 2001 Published: August 1, 2002
US20030054550
Cardiac hypertrophy factor and uses therefor
Inventors: Baker, Joffre; Chien, Kenneth;; King, Kathleen; Pennica, Diane; Wood, William;
Application No. 107931 Filed: March 26, 2002 Published: March 20, 2003
US 20030068678
WISP Polypeptides and nucleic acids encoding the same
Inventors): Levine, Arnold, J.; Pennica, Diane
Application No. 1 12267, Filed March 27, 2002, A2 Published April 10, 2003
Patent Number: 6642024
Issued November 4, 2003
Guanylate-Binding Protein
Inventor: Diane Pennica
ABSTRACTS
1 . Walsh, M.L., Pennica, D. and Cohen, P.S. Location of Specific mRN As on
Polysomes. Fed. Proc. 34, 637 (1975).
2. Ennis, H.L., Walsh, M.L., Walker, A.C., Pennica, D. and Cohen, P.S. Synthesis
and Decay of mRNA: Role of Ribosome-mRNA Interactions. Fed. Proc. 35.
1440(1976).
3. Pennica, D. and Cohen, P.S. Regulation of Ribosome Function during T4
Infection. Annual Meeting of American Society for Microbiology, 194 (1977).
4. Pennica, D., Lynch, K.R., Cohen, P.S. and Ennis, H.L. Functional Decay of
Vesicular Stomatitis Virus (VSV) mRNA In Vivo. Annual Meeting of American
Society of Microbiology, 244 (1979).
Holloway, B.P., Pennica, D., Heyward, J.T. and Obijeski, J.F. Rabies Virus
Transcription and Translation. Annual Meeting of American Society for
Microbiology (1980).
6. Pennica, D. Biosynthesis and Structure of Human Tissue-type Plasminogen
Activator. IXth International Congress on Thrombosis and Haemostasis (1983);
Stockholm, Sweden.
Levine, A.J., Reich, N., Pennica, D. and Goeddel, D.V. The Regulation of
Cellular Gene Expression in Transformed Cells. Modification of Cell Growth
Regulation by Oncogenic Products (1983); Annapolis, Maryland.
8. Pennica, D. Recent Progress in the Cloning of Human Tissue Plasminogen
Activator Gene. Sixth International Congress on Fibrinolysis, 65 (1985);
Lausanne, Switzerland.
9. Aggarwal, B.B., Hass, P.E., Kohr, W.J., Nedwin, GE. Gray, P.W., Pennica, D.,
Svedersky, L.P., Lee, S.-H., Palladino, M.A. and Goeddel, D.V. Biochemical and
Biological Properties of Human Lymphotoxin and Tumor Necrosis Factor. 14th
International Congress of Chemotherapy (1985).
10. Nedwin, GE., Pennica, D., Jarrett, J.A., Smith, D., Gray, P.W. and Goeddel,
D.V. The Human Lymphotoxin Gene: Structure and Homology with Tumor
Necrosis Factor. 1985 UCLA Symposia: Perspectives in Inflammation,
Neoplasia and Vascular Cell Biology.
11. Aggarwal, B.B., Pennica, D., Gray, P.W. and Goeddel, D.V. Biochemistry and
Molecular Biology of TNF and Lymphotoxin. CIBA Foundation, January 198
12. Pennica, D., King, K.L., Chien, K. R., Baker, J. B., and Wood, W. I.
Cardiotrophin-1, a Novel Cytokine that Induces Cardiac Myocyte Hypertrophy.
Cytokine, 6 No. 5, p. 577, 1994
13. Francis, N., Habecker, B., Pennica, D., and Landis, S. The Sweat Gland Derived
Cholinergic Differentiation Factor is Distinct From LIF, CNTF and CT-1. The
Society for Neuroscience, Nov. 11-16, 1995.
14. Cheng, J-G, Pennica, D., and Patterson, P. Cardiotrophin-1 Induces the Same
Neuropeptides in Sympathetic Neurons as the Neuropoietic Cytokines. The
Society for Neuroscience, San Diego, CA, Nov. 11-16, 1995
15. Sheng, Z., Pennica, D., Wood, W. I., Chien, K. R. Cardiotrophin-1 (CT-1): a
gp!30 Dependent Cytokine Promotes Cardiac Myocyte Survival and Displays
85
Restricted Expression to the Myocardium in the Early Murine Embryonic Heart.
American Heart Association Meeting, Anaheim, CA, Nov. 13-16, 1995
16. Wollert, K. C., Pennica, D., Paoni, N. F., Wood, W. 1., and Chien, K. R.
Divergent Signaling Pathways Mediate the Induction of the Atrial Natriuretic
Factor Gene During Cardiotrophin-1 Versus a-Adrenergic Receptor Mediated
Cardiomyocyte Hypertrophy. German Society of Cardiology, Mannheim,
Germany, April, 1995
17. Wollert, K. C., Pennica, D., Paoni, N. F., Glembotski, C. C., Wood, W. I., Taga,
T., Kishimoto, T. and Chien, K. R. Divergent Signaling Pathways Mediate the
Induction of the Atrial Natriuretic Factor Gene During Cardiotrophin-1 Versus
Phenylephrine Mediated Cardiomyocyte Hypertrophy. American 1 leart
Association Meeting, Anaheim, CA, Nov. 13-16, 1995
18. Yang. R., Jin, H., Pennica, D., Wood, W. I., and Paoni, N. F. Hemodynamic
Effects of Cardiotrophin-1, A New Cytokine, in Conscious Rats American Heart
Association Meeting, Anaheim, CA, Nov. 13-16, 1995
19. Jin, H., Yang, R., Li, W., Lai, J., Bennett, G, Nuijens, A., Ko, A., Keller, G,
Ryan, A., Pennica, D., Wood, W. I. and Paoni, N. F. Analysis of the Role of
Cardiotrophin-1 on Cardiac Hypertrophy In Vivo American Heart Association
Meeting, Anaheim, CA, Nov. 13-16, 1995
20. Kabakoff, S. L., Pennica, D., Chin, Y.E., Fu, X-Y, and Horowitz, M. C.
Cardiotrophin-1 : A New Autocrine Factor for Osteoblasts with Activities Distinct
from L1F American Society for Bone and Mineral Research, Seattle, Washington,
Sept. 7-11, 1996
21. Henderson, C.E., Arce, V, Swanson, T. A., Vejsada R., Pollock, R.A., Armanini,
M., Dudley, K., Phillips, H., Rosenthal, A., Kato, A., and Pennica, D.
Cardiotrophin-1 (CT-1), a Cytokine Secreted by Embryonic Muscle, Supports
Long-Term Survival of a Sub-population of Motoneurons Society of
Neuroscience, Washington, D.C., November 16-21, 1996
22. Spencer, S., Pennica, D., Gibbs, V.C., and Aguet, M. The Orphan Class II
Cytokine Receptor CRF2-4 International Society for Interferon and Cytokine
Research Geneva, Switzerland, October 6- 10, 1996
23. Gard, A. L., Pennica, D., Moore, A., and Ford 111 O.H. Cardiotrophin-1 Induces
an Astrocyte Phenotype in Cultured Oligodendrocyte Progenitors Society of
Neuroscience 1997
24. H. Mitsumoto, Klinkosz, B., Ishiyama, T., Tsuzaka, K. and Pennica, D.
Cardiotrophin- 1 has Marked Protective Effects in Wobbler Mouse Motor Meuron
Disease Society of Neuroscience 1997
25. Padrun, V., Hornfeld, D., Deglon, N., Pennica, D., Aebischer, P., and Zum, A.D.
Combined Effiects of Neurotrophins and Cytokines: Which are the Best
Neurotrophic Factor Combinations for Motoneurons? Society of Neuroscience
1997
26. Tsuzaka, K., Klinlosz, T., Ishiyama, E. P., Pioro, H., Mitsumoto and Pennica, D.,
Cardiotrophin- 1 has Marked Neuroprotective Effects in Wobbler Mouse Motor
Neuron Disease Society of Neuroscience 1997
27. Bysani Chandrasekar, Peter C Melby, Diane Pennica, and Gregory L. Freeman.
Expression of cardiotrophin- 1 and gp!30 during experimental acute Chagasic
cardiomyopathy. Circulation. 96(8): 1-424 (2371 A), 1997)
28. Taneyhill, L. A., Pennica, D., and Levine, A. J. Identification of Downstream
Genes in the Wnt-1 Signal Transduction Pathway Wnt Meeting, Cambridge,
1998
29. Gibbs, V. C., Swanson, T. A., Hillan, K. J., Keller, G-A., and Pennica, D., GBP-
4: A New Guanylate Binding Protein Expressed in Gastric Cancer Molecular
Mechanisms for Gastrointestinal Cancer Keystone Symposium April 1999
30. Lifeng Xu, Ryan Corcoran, James W. Welsh, Diane Pennica, Arnold Levine
Characterization of WISP- 1 : A Downstream Target of b-Catenin that is
Potentially Involved in Tumorigenesis Wnt Meeting, Stanford, July 1999
3 1 . Roux, F.J., He, S., Chupp, G, Mahboubi, K., Pennica, D., Panettieri, R. A.,
Pober, J., and Elias, J.A. Cardiotrophin- 1 (CT-1) Induces Human Airway Smooth
Muscle (HASM) Proliferation American Thoracic Society Meeting, Toronto,
Canada May, 2000
32. Chupp, GL., Roux, F.J., Homer, R.J., Redlich, C., Pennica, D., and Elias, J.A.
Immunohistochemical Localization of Cardiotrophin- 1 (CT-1) in Normal Human
Lung, American Thoracic Society Meeting, Toronto, Canada May, 2000
87 APPHNDIX B
lOWAM) U KNHCOT. UASSACMUSrni. CMAWMAN
CLAO*M n U. KHOM ISLAND OMMM HATCH UTAH
MOWAAO U MCT2INIAUU. OHK> NANCY LANOON KAStlBAUM KAN&AS
trAAX U UATSUNAOA. HAWAII JW MltOfOt. Vf MIOHT
CMMf TOTMtH J 0000. COMNICTICUT DAN COATS. MOMMA
MIA SIMON. ILMOIS STROM TMUKMOND. SOUTH CAKOUNA
TOM HAMM. IOWA OAVI DUOINStftGI H UINNISOTA
OC AOAAIS WASMNOTON TMAO COCHWN. UKSISSI fl
ANIAIU A MIKIASKI
Bnitefl States Senate
UTTUmiD. STA> OUWCTOII AND CMIf COUWSIl ^.....^^ ~*.
MSTIM A rvtutoN UWOMTV sTwf cxwcToii COMMITTEE ON LABOR AND
HUMAN RESOURCES
WASHINGTON. DC 20510-6300
April 13, 1989
Dr. Diane Pennica
Genentech, Incorporated
460 Point San Bueno Boulevard
South San Francisco, California 94080
Dear Dr. Pennica:
I have just learned that you are to receive the "Inventor of
the Year" award from Intellectual Property Owners Foundation for
dramatically facilitating the treatment of heart attack and
stroke victims.
The isolation and cloning of tissue plasminogen activator is
a remarkable scientific achievement, indeed, for which you should
be justly proud. I am sure the availability of TPA will, in and
of itself, offer new hope to victims of heart attacks and
strokes. Equally important, however, is the basic knowledge
gained through the insight, diligence, and expertise of
individuals such as yourself. You are to be commended for your
role in this pioneering enterprise.
Once again, congratulations and best wishes.
With warm regards,
Ever sincerely,
Claiborne Pell
Chairman
Subcommittee on Education,
Arts and Humanities
88
89
^L, !
David V. Goeddel, Diane Pennica, William J. Kohr, Gordon A. Kehar,
recipients of Inventor of the Year Award, 1989
Photo courtesy of Diane Pennica
91
APPENDIX C
Photos courtesy of Diane Pennica
USIKQ
OF CLONE
3a p- w-E-y-c-o PROBE
(2345
u 1 9 9 f l ^"
t * *
**-***
Photograph of the film on notebook page dated September 7, 1981, showing the
Colony Hybridization of 96 of the 5,000 clones that we screened with the
32P-TC(A/G)CA(A/G)TA(C/T)TCCCA(WEYCD) probe.
Clone 25E10 was one of the many clones we chose to sequence based on a
slightly darker signal with the probe than other clones.
-DP
92
1
TITLE
Mdanev V>M v -.0! Projecl No. JL__ ^
QJSUsi : - . Booh No 63
1
From P*
i
.
fl No
Ocx 20 1 1: >7 1 i
v, , : Ar^> J^f oa_ ( l |c n^i ^t>| /C 1 KJO^U
.. C^ It.ar-i i** l^-jw**fr5
f 7. 5 1
1 10
ra Ar jr. jly alu Arj fib* lu ey aly ii il 1 J tl
HO TCb CC.C .1.1 ,iS CCG TIC CTG T6C iCO ICC *A CTC ATC 4CC
Z3 M n c -pfc* S(10 J" <^ lOor^.
tCC TCC TOG ATT CT
etc CCC AAi- CTi l<
: TCT .cci ccc C.T 6<T_jtec ACC ACA <ICT -MC
40
J I J 11C ITC : ACA ACA TAC CSC CTC C1
r^n 1
5 J
oro gly jij jlj .la ill iy of* olu < 1 i 1 u lyi tyr 11* vl
CCT GCC 4*1 &Ai CAG CA . 4A4 riT SAA -ilC GAA 4AA I4C 4TT .TC
71
CAT 4i 1AA TIC >A
JU..J-I.
T OAT vC ACT IAC CAC 4AT 6AC ATT ,6CJ_ C,1<1
aln
fA -^ R. -.M 7?bp Rsf pteae
1 Ifl
cr* r i pro * r ; .so *f- ! or tro -,* tyr .l
. *C MA AAC CCA CAT CA * TCyA JIAC C<C TC<i TCC TAC %f C
F* *^t **
TTT AAO iC- C-6% AAi 1AC ACC. 1C A fi/i TIC ICC Aft,
T*i K Pr4 i-^r
>^>. -^ * *
fr.n,l.t.d -.I. ,
> io" i ?^S7.tO
ro K9 No
Wttnft*d & Understood by m.
Date Ilnv*n1d by . Oat*
1 ^^ f / < 1
One of the most exciting moments in the t-PA cloning project happened on October 20,
1981, when we determined that clone 25E10 contained a portion of the t-PA cDNA. This
notebook page shows the first bit of sequence from clone 25E10 that told us we had at last
isolated a t-PA cDNA clone. Only one other clone out of the 5000, Clone 25B2, contained
a partial t-PA cDNA sequence and it was on the same filter, but we did not choose to
sequence it initially because it did not stand out from the clones surrounding it.
The thrill of this was that 36 years after t-PA was discovered as a fibronolytic agent in
blood, we determined its structure by cloning.
-DP
93
III*
$c v
.; D> ^. IF, V , . , - . .- I- -. . - >-. , , .5i , JC .. ,, A. , . , - , ; -
> j E < v i-
- . . - . v5-ec*<r-.
CG
* Umtotslood by m.
Invwnludby
Entry from my notebook dated 11/11/81 showing the entire t-PA cDNA
sequence from Clone 25E10. Unfortunately, this clone was missing a small
number of amino acids from the 5 prime end of the protein. It took us another
five months of non-stop work 7 days a week and 14-16-hour days to get the
remainder of the t-PA cDNA clone.
-DP
APPENDIX D
Powerpoint slides courtesy of Diane Pennica
Recombinant t-PA Restores Blood Flow in Blocked Artery
Before t-PA After t-PA
Human Patient An giog ram in the Clinical Trial
Fibrin in a Blood Clot
Being
Dissolved by t-PA
95
APPENDIX E
Powerpoinl slides courtesy of Diane Pennicu
November 13, 1987
TIME IS OF THE ESSENCE
i MM nnM>( * kR rack |
40 Years After t-PA was First Identified
It was Approved As a Drug For
Heart Attack Patients
Ownted his tnougliTPAonluDdlomMl initial demand
Mountain
FDA approves heart drug
Medone diwct^s noost potonMy lawl ctoW/2
lot-dissolving drug called
step In coronary therapy
I B)ood-clo di*soh ing drug JF ^^ .^.^ ,
[wib.Hvm.JM.mrDA
From: Michael May
Date: October 7, 2004 7 OS 06 AM PDT
Subject: Please Forward to Ms. Diane Pennica, Chief Scientist
Catching Up From 191 J
Dear Ms Pennica
I was pleased to see/hear that you are still with Genentech And. like so many others I m sure, pleased that God made you
available to me on that dreadful day of December 17th. 1967
My wife had just had a post-op pulmonary embolism that stopped her heart They had already pronounced her dead when the ER
physician at Piano General Hospital gave her a new drug known as TPA We were both 33 years old and our girls were 13 and 9
Today, we are both SO and happily marned after 31 years and the girls are X and 26 both marned and college educated
Thank you so much for your work in helping others through science
Kind Regards.
Michael J May
Steve Birnbaum: First Heart Attack Patient Treated with t-PA
Christmas Card From Heart Attack Victim Mike Blum
97
Francis Wagner Stroke Survivor
Email from the Daughter of a Stroke Patient
My father had a massive stroke a few years ago. He was completely
paralyzed on one side, didn t know who he was. . .
They got him to the hospital very quickly, and an astute ER doctor
identified him as a candidate for t-PA therapy
10 minutes after the t-PA went in, he was waving, smiling and speaking fine.
He is now home, strolling around the neighborhood and showing
No obvious symptoms at all.
The doctors told my mother and brothers that they had never
seen such an amazing recovery. . .
So, needless to say, my family has a newfound appreciation
for the impact (and the future potential) of biotechnology.
98
INDEX-Diane Pennica
Aggarwal, Bart 29
APC (adenomatous polyposis coli) 52
B
Berg, Paul 62
Biogen 17
Bimbaum, Steve 27
Blum, Mike 26
Boyer, Herb 1 5
cardiac hypertrophy 45-46
cardiac myocyte hypertrophic factor
SeeCT-l
cardiotropin 1
SeeCJ-l
Chiron 8, 9, 10
Cohen, Paul 5, 7, 15
Col len. Desire 13-14, 15, 16,63
colon cancer 52, 54
CT-1 45-50,52,60
cloning process 48
D
Derynck, Rick 29, 30
E
education
family attitude about 2
paying for college 2
postdoc research at Roche Institute 7-10
SUNY-Fredonia 2-4
University of Rhode Island 5-6
embryonic stem cells 47
Ennis, Herb 7, 10
F
family
attitude about education 2
background 12, 15,26
marriage 26
Fox, Kevin 3-4, 25
G
gamma interferon 1 7, 30
Genentech
acquisition by Roche 58
cloning process 37-39
competitors 36
first impressions 10
key to getting job there 8
mammalian cell expression 22
management environment 58, 59
research environment 24-25, 26, 30-32, 35,
36, 39-40, 42, 56
technology used at 56
the company s legacy 60
See also specific research projects
Goeddel, Dave 9, 1 1 , 1 7, 1 9, 2 1 , 29, 30, 36, 4 1 , 42,
59, 62, 63
Griinenthal 11, 12, 16
H
Hayflick, Joel 29
Heyneker, Herb 9, 10, 1 1, 16, 17, 18
HGH (human growth hormone) 60
Hoffmann-La Roche
See Roche Institute of Molecular Biology
Holmes, Bill 12, 17,25
1
Ingelheim, Behringer 62
Innis, Mike 9
Kabi 19,20,21,36
Kiley, Tom 33
King, Kathy 47, 48, 56
Kleid, Dennis 1 1
Kohr, Bill 19,21,29,36,42,63
Levine, Arnie 34, 36, 41, 52, 59
Levinson, Art 34, 58
LIF (leukemia inhibitory factor) 47, 49
M
Marietta, Michael 25
N
Nedwin, Glenn 29
Nile, Hugh 10
O
Obijeski, Jack 8
Ochoa, Severe 10
Oren, Moshe 35, 36
100
p53 34-37, 41
Palladino, Michael 29
patent process 5 1
patents 61
litigation 62-63
publication 7, 20-21, 31-32, 35, 39, 50, 51, 55, 61
Raab, Kirk 58
Raines, Steve 62
Roche Institute of Molecular Biology 7-10
Ross, Mike 8, 10
Schmoyer, Irv 34
science
collaboration in 35, 45, 50
enthusiasm for 1 5
ethical concerns 59-60
good working strategy 59
overcoming disappointment 52
personal research style 57
personal rewards of 26-28, 30, 32, 40, 41, 48,
55,63
rivalries in 17, 18, 20-21, 22, 30, 32
self-identification as a scientist 61
teamwork in 56
value of persistence 37, 43, 57
value of to society 60
women in 15, 2425, 26
Seeburg, Peter 9, 10, 19, 29, 42
Stark, George 62
SUNY-Fredonia
accolades from 27
education at 24
suppressive subtractive hybridization 53
Swanson, Bob 10, 1 1, 16, 20, 30, 58
learning of its existence 12-16
patent litigation 62-63
patent process 32-34
research strategy 29
tandem research on urokinase 16-17
the breakthrough 1 9
the researcher group at Genentech 29
tumor necrosis factor
SeeTNF
U
Ullrich, Axel 10,42
urokinase 11-12, 16-17
uromodulin 4243
Varmus, Harold 53
Vehar, Gordon 12, 20, 21, 42, 63
W
Watson, James 62
Weissmann, Charles 17, 30
WISP 52-55, 56
cloning process 5354
Wnt-induced secreted protein
See WISP
Wood, William 46
Testa, Doug 7, 8
tissue plasminogen activator
See t-PA
TNF 29-3 1,38
announcement of clone 32
beginning the project 24
research strategy 29
t-PA 18-21,38,60,63
announcement of clone 2223
clinical application of 26-28
SALLY SMITH HUGHES
Sally Smith Hughes is a historian of science at ROHO whose research focuses on the recent
history of bioscience. She began work in oral history at the Bancroft Library in 1978 and joined
ROHO in 1980. She has conducted interviews for over 100 oral histories, whose subjects range
from the AIDS epidemic to medical physics. Her focus for the past decade has been on the
biotechnology industry in northern California. She is the author of The Virus: A History of the
Concept and an article in /.w .v, the journal of the History of Science Society, on the
commercialization of molecular biology.
Ji BERKELEY LIBRARIES
067700554
Open Library
